Notebook
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07/10
PCR
Yoshimoto
lane | Primer Foward | Primer Revese | Template |
---|
1 | shortAK2 | shortAK2 | B.xylophilus cDNA |
2 | Top1 | Top1 | B.xylophilus cDNA |
3 | long AK2 | long AK2 | B.xylophilus cDNA |
4 | eef-1g | eef-1g | B.xylophilus cDNA |
5 | AK1 | AK1 | B.xylophilus cDNA |
6 | Asb | Asb | B.xylophilus cDNA |
7 | 14-3-3zeta | 14-3-3zeta | B.xylophilus cDNA |
8 | tropomiosin | tropomiosin | B.xylophilus cDNA |
9 | 14-3-3 protein | 14-3-3 protein | B.xylophilus cDNA |
PCR Polymerase : 5*Q5 Polymerase
98.0°C : 30sec
98.0°C : 10sec
58.0°C : 30sec
72.0°C : 1min
72.0°C : 1min
4.0°C : Infinate
X35 cycle
PCR pulification: lane 2~9
Gel extraction: lane 2~9
07/11
Miniprep_Nanodrop
Yoshimoto
sample | Sample name | 260/280 | 260/230 | ng/ul |
---|
1 | 2 | no data | no data | 237.8 |
2 | 3 | no data | no data | 180.5 |
3 | 4 | no data | no data | 185.6 |
4 | 5 | no data | no data | 167.2 |
5 | 6 | no data | no data | 328.2 |
6 | 7 | no data | no data | 133.7 |
7 | 8 | no data | no data | 168.4 |
8 | 9 | no data | no data | 242.5 |
The way of extraction : Gel extraction
07/11
Ligation
Yoshimoto
sample | Vector name | Vector amount(ul) | Insert name | Insert amount(ul) |
---|
1 | YEplac195/sma1 | 1 | Template2 | 1.0 |
2 | YEplac195/sma1 | 1 | Template3 | 1.0 |
3 | YEplac195/sma1 | 1 | Template4 | 1.0 |
4 | YEplac195/sma1 | 1 | Template5 | 1.0 |
5 | YEplac195/sma1 | 1 | Template6 | 1.0 |
6 | YEplac195/sma1 | 1 | Template7 | 1.0 |
7 | YEplac195/sma1 | 1 | Template8 | 1.0 |
8 | YEplac195/sma1 | 1 | Template9 | 1.0 |
07/11
Transformation
Yoshimoto
sample | CompetentCell name | DNA name |
---|
1 | DH5a | 7/11 ligation product |
07/12
Ligation
Yoshimoto
sample | Vector name | Vector amount(ul) | Insert name | Insert amount(ul) |
---|
1 | pRS315 | 1 | Template | 1.0 |
07/12
Transformation
Yoshimoto
sample | CompetentCell name | DNA name |
---|
1 | DH5a | 7/12 ligation product 2 |
2 | DH5a | 7/12 ligation product 3 |
3 | DH5a | 7/12 ligation product 4 |
4 | DH5a | 7/12 ligation product 5 |
5 | DH5a | 7/12 ligation product 6 |
6 | DH5a | 7/12 ligation product 7 |
7 | DH5a | 7/12 ligation product 8 |
8 | DH5a | 7/12 ligation product 9 |
07/13
PCR
Yoshimoto
lane | Primer Foward | Primer Revese | Template |
---|
1 | T7Top1 | T7Top1 | 7/10 gel extraction product2 |
2 | T7long AK2 | T7long AK2 | 7/10 gel extraction product3 |
3 | T7eef-1g | T7eef-1g | 7/10 gel extraction product4 |
4 | T7AK1 | T7AK1 | 7/10 gel extraction product5 |
5 | T7Asb | T7Asb | 7/10 gel extraction product6 |
6 | T714-3-3zeta | T714-3-3zeta | 7/10 gel extraction product7 |
7 | T7tropomiosin | T7tropomiosin | 7/10 gel extraction product8 |
8 | T714-3-3 protein | T714-3-3 protein | 7/10 gel extraction product9 |
PCR Polymerase : ExTaq
98.0°C : 1min
98.0°C : 10sec
58.0°C : 30sec
72.0°C : 1min
72.0°C : 1min
4.0°C : Infinate
X35 cycle
07/14
Miniprep_Nanodrop
Yoshimoto
sample | Sample name | 260/280 | 260/230 | ng/ul |
---|
1 | 1 | 1.84 | 1.96 | 98.4 |
2 | 2 | 1.79 | 1.85 | 76.2 |
3 | 3 | 1.85 | 2.1 | 102.1 |
4 | 4 | 1.62 | 0.49 | 9.5 |
5 | 5 | 1.91 | 1.91 | 83.6 |
6 | 6 | 1.82 | 1.73 | 48.0 |
7 | 7 | 1.85 | 1.85 | 83.0 |
8 | 8 | 1.86 | 2.84 | 81.5 |
9 | 9 | 1.57 | 0.33 | 22.6 |
10 | 10 | 1.54 | 0.55 | 17.8 |
11 | 11 | 1.83 | 2.02 | 88.0 |
12 | 12.0 | 1.85 | 2.94 | 82.0 |
13 | 13.0 | 1.83 | 1.83 | 90.6 |
14 | 14.0 | 1.87 | 1.65 | 46.2 |
15 | 15.0 | 1.89 | 2.0 | 75.8 |
16 | 16.0 | 1.67 | 1.98 | 114.0 |
17 | 17.0 | 1.88 | 1.46 | 88.5 |
18 | 18.0 | 1.85 | 1.91 | 99.4 |
19 | 19.0 | 1.79 | 1.79 | 53.8 |
20 | 20.0 | 1.37 | 0.58 | 17.0 |
21 | 21.0 | 1.63 | 0.57 | 0.57 |
22 | 22.0 | 1.83 | 1.84 | 53.0 |
The way of extraction : promega
07/15
Restriction Enzyme digestion
Yoshimoto
lane | Sample DNA name | DNA amount (ul) | RE1 name | RE1 amount(ul) | RE2 name | RE2 amount(ul) | Buffer name | Buffer amount(ul) | Total volume(ul) |
---|
1 | 1 | 5.0 | Pst1 | 0.05 | Sac1 | 0.05 | 10xM | 1.0 | 10.0 |
2 | 2 | 5.0 | Pst1 | 0.05 | Sac1 | 0.05 | 10xM | 1.0 | 10.0 |
3 | 3 | 5.0 | Pst1 | 0.05 | Sac1 | 0.05 | 10xM | 1.0 | 10.0 |
4 | 4 | 5.0 | Pst1 | 0.05 | Sac1 | 0.05 | 10xM | 1.0 | 10.0 |
5 | 5 | 5.0 | Pst1 | 0.05 | Sac1 | 0.05 | 10xM | 1.0 | 10.0 |
6 | 6 | 5.0 | Pst1 | 0.05 | Sac1 | 0.05 | 10xM | 1.0 | 10.0 |
7 | 7 | 5.0 | Pst1 | 0.05 | Sac1 | 0.05 | 10xM | 1.0 | 10.0 |
8 | 8 | 5.0 | Pst1 | 0.05 | Sac1 | 0.05 | 10xM | 1.0 | 10.0 |
9 | 9 | 5.0 | Pst1 | 0.05 | Sac1 | 0.05 | 10xM | 1.0 | 10.0 |
10 | 10 | 5.0 | Pst1 | 0.05 | Sac1 | 0.05 | 10xM | 1.0 | 10.0 |
11 | 11 | 5.0 | Pst1 | 0.05 | Sac1 | 0.05 | 10xM | 1.0 | 10.0 |
12 | 12 | 5.0 | Pst1 | 0.05 | Sac1 | 0.05 | 10xM | 1.0 | 10.0 |
13 | 13 | 5.0 | Pst1 | 0.05 | Sac1 | 0.05 | 10xM | 1.0 | 10.0 |
14 | 14 | 5.0 | Pst1 | 0.05 | Sac1 | 0.05 | 10xM | 1.0 | 10.0 |
15 | 15 | 5.0 | Pst1 | 0.05 | Sac1 | 0.05 | 10xM | 1.0 | 10.0 |
16 | 16 | 5.0 | Pst1 | 0.05 | Sac1 | 0.05 | 10xM | 1.0 | 10.0 |
17 | 17 | 5.0 | Pst1 | 0.05 | Sac1 | 0.05 | 10xM | 1.0 | 10.0 |
18 | 18 | 5.0 | Pst1 | 0.05 | Sac1 | 0.05 | 10xM | 1.0 | 10.0 |
19 | 19 | 5.0 | Pst1 | 0.05 | Sac1 | 0.05 | 10xM | 1.0 | 10.0 |
20 | 20 | 5.0 | Pst1 | 0.05 | Sac1 | 0.05 | 10xM | 1.0 | 10.0 |
21 | 21 | 5.0 | Pst1 | 0.05 | Sac1 | 0.05 | 10xM | 1.0 | 10.0 |
22 | 22 | 5.0 | Pst1 | 0.05 | Sac1 | 0.05 | 10xM | 1.0 | 10.0 |
07/16
Miniprep_Nanodrop
Yoshimoto
sample | Sample name | 260/280 | 260/230 | ng/ul |
---|
1 | T7 3 | 1.86 | 2.28 | 255.7 |
2 | T7 4 | 1.83 | 2.13 | 121.9 |
3 | T7 5 | 1.83 | 2.29 | 224.2 |
4 | T7 6 | 1.85 | 2.18 | 146.1 |
5 | T7 7 | 1.85 | 2.25 | 96.5 |
6 | T7 8 | 1.85 | 2.27 | 165.4 |
7 | T7 9 | No data | No data | No data |
The way of extraction : promega
07/16
PCR
Yoshimoto
lane | Primer Foward | Primer Revese | Template |
---|
1 | short AK2 | short AK2 | B.xylophilus cDNA |
2 | Top1 | Top1 | B.xylophilus cDNA |
PCR Polymerase : Extaq
94.0°C : 1min
94.0°C : 30sec
60.0°C : 1min
72.0°C : 2min
72.0°C : 5min
4.0°C : Infinate
X35 cycle
08/06
Nebuilder
Yoshimoto
Sample | Vectorname | Vector amount (ul) | Insert1 name | Insert1 amount(ul) | Insert2 name | Insert2 amount (ul) | Mastermix amount (ul) | Total amount(ul) |
---|
1 | YEplac195/sma1 | 2.5 | Gal-AK | 0.8 | Ter-AK | 1.0 | 10.0 | 20.0 |
2 | YEplac195/sma1 | 2.5 | GPD-AK | 1.2 | Ter-AK | 1.0 | 10.0 | 20.0 |
08/06
Transformation
Yoshimoto
sample | CompetentCell name | DNA name |
---|
1 | DH5a | Insert |
08/07
Miniprep_Nanodrop
Yoshimoto
sample | Sample name | 260/280 | 260/230 | ng/ul |
---|
1 | 1 | 1.97 | 2.03 | 36.0 |
2 | 2 | 2.03 | 1.89 | 28.7 |
3 | 3 | 1.92 | 1.98 | 34.3 |
4 | 4 | 1.96 | 1.88 | 35.5 |
5 | 5 | 2.03 | 1.87 | 16.2 |
6 | 6 | 2.01 | 2.11 | 50.0 |
7 | 7 | 1.95 | 1.95 | 30.3 |
8 | 8 | 1.95 | 2.06 | 38.2 |
9 | 9 | 2.12 | 1.94 | 17.0 |
10 | 10 | no data | no data | no data |
11 | 11 | 1.99 | 1.92 | 27.9 |
12 | 12 | 1.96 | 1.79 | 19.1 |
13 | 13 | 1.95 | 1.96 | 29.4 |
14 | 14 | 1.98 | 1.64 | 22.7 |
The way of extraction : fastgene plasmid mini kit
08/08
Transformation
Yoshimoto
sample | CompetentCell name | DNA name |
---|
1 | DH5a | 8/6 Nebuilder product |
2 | DH5a | 8/6 Nebuilder product |
3 | DH5a | 8/6 Nebuilder product |
4 | DH5a | 8/6 Nebuilder product |
5 | DH5a | 8/6 Nebuilder product |
6 | DH5a | 8/6 Nebuilder product |
7 | DH5a | 8/6 Nebuilder product |
8 | DH5a | 8/6 Nebuilder product |
08/08
Restriction Enzyme digestion
Yoshimoto
lane | Sample DNA name | DNA amount (ul) | RE1 name | RE1 amount(ul) | Buffer name | Buffer amount(ul) | Total volume(ul) |
---|
1 | miniprep 1 | 5.0 | Pst1 | 0.05 | 10xH | 1.0 | 10.0 |
2 | miniprep 2 | 5.0 | Pst1 | 0.05 | 10xH | 1.0 | 10.0 |
3 | miniprep 3 | 5.0 | Pst1 | 0.05 | 10xH | 1.0 | 10.0 |
4 | miniprep 4 | 5.0 | Pst1 | 0.05 | 10xH | 1.0 | 10.0 |
5 | miniprep 5 | 5.0 | Pst1 | 0.05 | 10xH | 1.0 | 10.0 |
6 | miniprep 7 | 5.0 | Pst1 | 0.05 | 10xH | 1.0 | 10.0 |
08/09
Miniprep_Nanodrop
Yoshimoto
sample | Sample name | 260/280 | 260/230 | ng/ul |
---|
1 | 8 | 1.69 | 1.32 | 12.1 |
2 | 9 | 1.97 | 1.45 | 17.0 |
3 | 11 | 1.83 | 0.99 | 12.5 |
4 | 12 | 1.92 | 1.05 | 12.3 |
The way of extraction : promega PureYield Plasmid Midiprepkit
08/09
Restriction Enzyme digestion
Yoshimoto
lane | Sample DNA name | DNA amount (ul) | RE1 name | RE1 amount(ul) | Buffer name | Buffer amount(ul) | Total volume(ul) |
---|
1 | 8/8 miniprep8 | 5.0 | Pst1 | 0.05 | 10xH | 1.0 | 10.0 |
2 | 8/8 miniprep9 | 5.0 | Pst1 | 0.05 | 10xH | 1.0 | 10.0 |
3 | 8/8 miniprep11 | 5.0 | Pst1 | 0.05 | 10xH | 1.0 | 10.0 |
4 | 8/8 miniprep12 | 5.0 | Pst1 | 0.05 | 10xH | 1.0 | 10.0 |
08/09
Restriction Enzyme digestion
Fukuda
lane | Sample DNA name | DNA amount (ul) | RE1 name | RE1 amount(ul) | RE2 name | RE2 amount(ul) | Buffer name | Buffer amount(ul) | Total volume(ul) |
---|
1 | AK2 | 5.0 | Xba1 | 0.5 | EcoR1 | 0.5 | 10xM | 6.0 | 36.0 |
2 | AK2 | 5.0 | Xba1 | 0.5 | EcoR1 | 0.5 | 10xM | 6.0 | 36.0 |
3 | eef-1g | 5.0 | Xba1 | 0.5 | EcoR1 | 0.5 | 10xM | 6.0 | 36.0 |
4 | eef-1g | 5.0 | Xba1 | 0.5 | EcoR1 | 0.5 | 10xM | 6.0 | 36.0 |
5 | AK1 | 5.0 | Xba1 | 0.5 | EcoR1 | 0.5 | 10xM | 6.0 | 36.0 |
6 | AK1 | 5.0 | Xba1 | 0.5 | EcoR1 | 0.5 | 10xM | 6.0 | 36.0 |
7 | Asb | 5.0 | Xba1 | 0.5 | EcoR1 | 0.5 | 10xM | 6.0 | 36.0 |
8 | Asb | 5.0 | Xba1 | 0.5 | EcoR1 | 0.5 | 10xM | 6.0 | 36.0 |
9 | 14-3-3-zeta | 5.0 | Xba1 | 0.5 | EcoR1 | 0.5 | 10xM | 6.0 | 36.0 |
10 | 14-3-3-zeta | 5.0 | Xba1 | 0.5 | EcoR1 | 0.5 | 10xM | 6.0 | 36.0 |
11 | tropomiosin | 5.0 | Xba1 | 0.5 | EcoR1 | 0.5 | 10xM | 6.0 | 36.0 |
12 | tropomiosin | 5.0 | Xba1 | 0.5 | EcoR1 | 0.5 | 10xM | 6.0 | 36.0 |
13 | 14-3-3 protein | 5.0 | Xba1 | 0.5 | EcoR1 | 0.5 | 10xM | 6.0 | 36.0 |
14 | 14-3-3 protein | 5.0 | Xba1 | 0.5 | EcoR1 | 0.5 | 10xM | 6.0 | 36.0 |
08/10
Nebuilder
Yoshimoto
Sample | Vectorname | Vector amount (ul) | Insert1 name | Insert1 amount(ul) | Insert2 name | Insert2 amount (ul) | Mastermix amount (ul) | Total amount(ul) |
---|
1 | YEplac195/sma1 | 2.5 | Gal-AK | 0.8 | Ter-AK | 1.0 | 10.0 | 20.0 |
2 | YEplac195/sma1 | 2.5 | GPD-AK | 1.2 | Ter-AK | 1.0 | 10.0 | 20.0 |
08/10
Transformation
Yoshimoto
sample | CompetentCell name | DNA name |
---|
1 | DH5a | insert |
08/10
PCR
Ohara
lane | Primer Foward | Primer Revese | Template |
---|
1 | T7AK2F | T7AK2R | B.xylophils cDNA1 |
2 | T7AK2F | T7AK2R | B.xylophils cDNA2 |
3 | T7Egf-1gF | T7Egf-1gR | B.xylophils cDNA3 |
4 | T7Egf-1gF | T7Egf-1gR | B.xylophils cDNA4 |
5 | T7AK1F | T7Ak1R | B.xylophils cDNA5 |
6 | T7AK1F | T7Ak1R | B.xylophils cDNA6 |
7 | T714-3-3zetaF | T714-3-3 zetaR | B.xylophils cDNA7 |
8 | T714-3-3zetaF | T714-3-3 zetaR | B.xylophils cDNA8 |
9 | T7tropomiosin F | T7tropomiosin R | B.xylophils cDNA9 |
10 | T7tropomiosin F | T7tropomiosin R | B.xylophils cDNA10 |
11 | T714-3-3 F | T714-3-3 R | B.xylophils cDNA11 |
12 | T714-3-3 F | T714-3-3 R | B.xylophils cDNA12 |
13 | T7AK2F | T7AK2R | water |
PCR Polymerase : gotaq
94.0°C : 2min
94.0°C : 30sec
62.0°C : 30sec
68.0°C : 1min20sec
68.0°C : 7min
4.0°C : Infinate
X30 cycle
08/11
colony_PCR
Yoshimoto
lane | Primer Foward | Primer Revese | Template |
---|
1 | M13F | M13R | 1 |
2 | M13F | M13R | 3 |
3 | M13F | M13R | 5 |
4 | M13F | M13R | 7 |
5 | M13F | M13R | 9 |
6 | M13F | M13R | 11 |
7 | M13F | M13R | 13 |
8 | M13F | M13R | 15 |
9 | M13F | M13R | 17 |
10 | M13F | M13R | 19 |
11 | M13F | M13R | 21 |
12 | M13F | M13R | 23 |
13 | M13F | M13R | 25 |
14 | M13F | M13R | 27 |
15 | M13F | M13R | 29 |
16 | M13F | M13R | 31 |
17 | M13F | M13R | 33 |
18 | M13F | M13R | 35 |
19 | M13F | M13R | 37 |
20 | M13F | M13R | 39 |
21 | M13F | M13R | 41 |
22 | M13F | M13R | 43 |
23 | M13F | M13R | 45 |
24 | M13F | M13R | 47 |
25 | M13F | M13R | 49 |
PCR Polymerase : QuickTaq
94.0°C : 2min
94.0°C : 30sec
60.0°C : 30sec
72.0°C : 1min
72.0°C : 1min
4.0°C : Infinate
X35 cycle
08/11
Restriction Enzyme digestion
Ohara
lane | Sample DNA name | DNA amount (ul) | RE1 name | RE1 amount(ul) | Buffer name | Buffer amount(ul) | Total volume(ul) |
---|
1 | 1-1 | 5.0 | Pst1 | 1.0 | 10xH | 1.0 | 3.0 |
2 | 1-2 | 5.0 | Pst1 | 1.0 | 10xH | 1.0 | 3.0 |
3 | 1-3 | 5.0 | Pst1 | 1.0 | 10xH | 1.0 | 3.0 |
4 | 1-4 | 5.0 | Pst1 | 1.0 | 10xH | 1.0 | 3.0 |
5 | 1-5 | 5.0 | Pst1 | 1.0 | 10xH | 1.0 | 3.0 |
6 | 2-1 | 5.0 | Pst1 | 1.0 | 10xH | 1.0 | 3.0 |
7 | 1-6 | 5.0 | Pst1 | 1.0 | 10xH | 1.0 | 3.0 |
8 | 2-2 | 5.0 | Pst1 | 1.0 | 10xH | 1.0 | 3.0 |
9 | 2-3 | 5.0 | Pst1 | 1.0 | 10xH | 1.0 | 3.0 |
10 | 2-4 | 5.0 | Pst1 | 1.0 | 10xH | 1.0 | 3.0 |
11 | 2-5 | 5.0 | Pst1 | 1.0 | 10xH | 1.0 | 3.0 |
12 | 2-6 | 5.0 | Pst1 | 1.0 | 10xH | 1.0 | 3.0 |
13 | 3-1 | 5.0 | Pst1 | 1.0 | 10xH | 1.0 | 3.0 |
14 | 3-2 | 5.0 | Pst1 | 1.0 | 10xH | 1.0 | 3.0 |
15 | 3-3 | 5.0 | Pst1 | 1.0 | 10xH | 1.0 | 3.0 |
16 | 3-4 | 5.0 | Pst1 | 1.0 | 10xH | 1.0 | 3.0 |
08/11
Miniprep_Nanodrop
Yoshimoto
sample | Sample name | 260/280 | 260/230 | ng/ul |
---|
1 | 1 | 1.88 | 1.6 | 116.8 |
2 | 8 | 1.9 | 1.67 | 170.0 |
3 | 9 | 1.91 | 1.71 | 167.6 |
4 | 12 | 1.88 | 1.6 | 135.6 |
The way of extraction : promega PureYield Plasmid Midiprepkit
08/11
colony_PCR
Yoshimoto
lane | Primer Foward | Primer Revese | Template |
---|
1 | M13F | M13R | 2 |
2 | M13F | M13R | 4 |
3 | M13F | M13R | 6 |
4 | M13F | M13R | 8 |
5 | M13F | M13R | 10 |
6 | M13F | M13R | 12 |
7 | M13F | M13R | 14 |
8 | M13F | M13R | 16 |
9 | M13F | M13R | 18 |
10 | M13F | M13R | 20 |
11 | M13F | M13R | 22 |
12 | M13F | M13R | 24 |
13 | M13F | M13R | 26 |
14 | M13F | M13R | 28 |
15 | M13F | M13R | 30 |
16 | M13F | M13R | 32 |
17 | M13F | M13R | 34 |
18 | M13F | M13R | 36 |
19 | M13F | M13R | 38 |
20 | M13F | M13R | 40 |
21 | M13F | M13R | 42 |
22 | M13F | M13R | 44 |
23 | M13F | M13R | 46 |
24 | M13F | M13R | 48 |
25 | M13F | M13R | 50 |
26 | M13F | M13R | 53 |
27 | M13F | M13R | 54 |
28 | M13F | M13R | 55 |
29 | M13F | M13R | 56 |
PCR Polymerase : Quick_taq
94.0°C : 2min
94.0°C : 30sec
60.0°C : 30sec
72.0°C : 1min
72.0°C : 10min
4.0°C : Infinate
X35 cycle
08/11
Ligation
Yoshimoto
sample | Vector name | Vector amount(ul) | Insert name | Insert amount(ul) |
---|
1 | YEplac195/sma1 | 1 | AK2-gal1 | 1.0 |
2 | YEplac195/sma1 | 1 | AK2-GPD | 1.0 |
08/11
Restriction Enzyme digestion
Ohara
lane | Sample DNA name | DNA amount (ul) | RE1 name | RE1 amount(ul) | Buffer name | Buffer amount(ul) | Total volume(ul) |
---|
1 | 7A | 5.0 | Pst1 | 0.5 | 10xH | 6.0 | 60.0 |
2 | 7B | 5.0 | Pst1 | 0.5 | 10xH | 6.0 | 60.0 |
08/11
PCR
Yoshimoto
lane | Primer Foward | Primer Revese | Template |
---|
1 | M13F | M13R | AK2-gal1 Nebuilder |
2 | M13F | M13R | AK2 GPD Nebuilder |
3 | M13F | M13R | gfp gal Nebuilder |
4 | M13F | M13R | gfp GPD Nebuilder |
5 | M13F | M13R | AK1 gal Nebuilder |
6 | M13F | M13R | AK1 GPD Nebuilder |
PCR Polymerase : 2*KAPA 2G Robust HS Readymix
94.0°C : 2min
94.0°C : 15sec
60.0°C : 15sec
72.0°C : 40sec
72.0°C : 1min
4.0°C : Infinate
X35 cycle
08/11
Transformation
Yoshimoto
sample | CompetentCell name | DNA name |
---|
1 | DH5a | insert |
08/12
PCR
Ohara
lane | Primer Foward | Primer Revese | Template |
---|
1 | AK1F | AK1R | 8 |
2 | AK1F | AK1R | 9 |
3 | AK1F | AK1R | 12 |
4 | AK1F | AK1R | 8/6 gal |
5 | AK1F | AK1R | 8/6 GPD |
6 | AK1F | AK1R | gal1 AK1 |
7 | AK1F | AK1R | gal1 AK1 |
8 | AK1F | AK1R | water |
PCR Polymerase : quicktaq
94.0°C : 2min
94.0°C : 30sec
60.0°C : 30sec
68.0°C : 1min
68.0°C : 7min
4.0°C : Infinate
X35 cycle
08/12
Ligation
Yoshimoto
sample | Vector name | Vector amount(ul) | Insert name | Insert amount(ul) |
---|
1 | YEplac195/sma1 | 1 | gblock 71 | 1.0 |
2 | YEplac195/sma1 | 1 | gblock 72 | 1.0 |
3 | YEplac195/sma1 | 1 | gblock 73 | 1.0 |
4 | YEplac195/sma1 | 1 | gblock 74 | 1.0 |
5 | YEplac195/sma1 | 1 | gblock 75 | 1.0 |
6 | YEplac195/sma1 | 1 | gblock 76 | 1.0 |
7 | YEplac195/sma1 | 1 | gblock 77 | 1.0 |
8 | YEplac195/sma1 | 1 | gblock 78 | 1.0 |
08/12
Transformation
Yoshimito
sample | CompetentCell name | DNA name |
---|
1 | DH5a | insert |
08/13
PCR
Ohara
lane | Primer Foward | Primer Revese | Template |
---|
1 | AK2F | AK2F | 8 |
2 | AK2F | AK2F | 9 |
3 | AK2F | AK2F | 12 |
4 | AK2F | AK2F | 8/6 gal |
5 | AK2F | AK2F | 8/6 GPD |
6 | AK2F | AK2F | 7 |
7 | AK2F | AK2F | water |
PCR Polymerase : 2*KAPA hifi HS ready mix
95.0°C : 3min
98.0°C : 20sec
60.0°C : 15sec
72.0°C : 2min
72.0°C : 2min
4.0°C : Infinate
X40 cycle
08/13
Restriction Enzyme digestion
Ohara
lane | Sample DNA name | DNA amount (ul) | RE1 name | RE1 amount(ul) | Buffer name | Buffer amount(ul) | Total volume(ul) |
---|
1 | 1 | 5.0 | Pst1 | 1.0 | 10xH | 1.0 | 10.0 |
2 | 2 | 5.0 | Pst1 | 1.0 | 10xH | 1.0 | 10.0 |
3 | 3 | 5.0 | Pst1 | 1.0 | 10xH | 1.0 | 10.0 |
4 | 4 | 5.0 | Pst1 | 1.0 | 10xH | 1.0 | 10.0 |
5 | 5 | 5.0 | Pst1 | 1.0 | 10xH | 1.0 | 10.0 |
6 | 7 | 5.0 | Pst1 | 1.0 | 10xH | 1.0 | 10.0 |
7 | 1 | 5.0 | Pst1 | 1.0 | 10xH | 1.0 | 10.0 |
8 | 7 | 5.0 | Pst1 | 1.0 | 10xH | 1.0 | 10.0 |
9 | 9 | 5.0 | Pst1 | 1.0 | 10xH | 1.0 | 10.0 |
10 | 8 | 5.0 | Pst1 | 1.0 | 10xH | 1.0 | 10.0 |
11 | 12 | 5.0 | Pst1 | 1.0 | 10xH | 1.0 | 10.0 |
08/13
Restriction Enzyme digestion
Ohara
lane | Sample DNA name | DNA amount (ul) | RE1 name | RE1 amount(ul) | Buffer name | Buffer amount(ul) | Total volume(ul) |
---|
1 | 14 | 5.0 | Pst1 | 1.0 | 10xH | 1.0 | 10.0 |
2 | 16 | 5.0 | Pst1 | 1.0 | 10xH | 1.0 | 10.0 |
3 | 22 | 5.0 | Pst1 | 1.0 | 10xH | 1.0 | 10.0 |
4 | 24 | 5.0 | Pst1 | 1.0 | 10xH | 1.0 | 10.0 |
5 | 26 | 5.0 | Pst1 | 1.0 | 10xH | 1.0 | 10.0 |
6 | 30 | 5.0 | Pst1 | 1.0 | 10xH | 1.0 | 10.0 |
7 | 36 | 5.0 | Pst1 | 1.0 | 10xH | 1.0 | 10.0 |
8 | 24 | 5.0 | Pst1 | 1.0 | 10xH | 1.0 | 10.0 |
9 | 14 | 5.0 | Pst1 | 0.0 | 10xH | 0.0 | 10.0 |
10 | 22 | 5.0 | Pst1 | 0.0 | 10xH | 0.0 | 10.0 |
11 | 36 | 5.0 | Pst1 | 0.0 | 10xH | 0.0 | 10.0 |
08/13
colony PCR
Ohara
lane | Primer Foward | Primer Revese | Template |
---|
1 | M13F | M13R | 77-1 |
2 | M13F | M13R | 77-2 |
3 | M13F | M13R | 77-3 |
4 | M13F | M13R | 78-4 |
5 | M13F | M13R | 78-5 |
PCR Polymerase : 2*KAPA hifi HS ready mix
95.0°C : 1min
98.0°C : 20sec
60.0°C : 15sec
72.0°C : 1min
72.0°C : 1min
4.0°C : Infinate
X35 cycle
08/13
Miniprep_Nanodrop
Yoshimoto
sample | Sample name | 260/280 | 260/230 | ng/ul |
---|
1 | 14 | 1.91 | 1.44 | 82.6 |
2 | 16 | 1.96 | 1.67 | 91.4 |
3 | 22 | 2.04 | 1.77 | 96.7 |
4 | 24 | 1.88 | 1.5 | 93.0 |
5 | 26 | 1.89 | 1.51 | 89.4 |
6 | 30 | 1.9 | 1.48 | 74.6 |
7 | 36 | 1.91 | 1.5 | 87.5 |
The way of extraction : promega PureYield Plasmid Miniprepkit
08/13
Ligation
Yoshimoto
sample | Vector name | Vector amount(ul) | Insert name | Insert amount(ul) |
---|
1 | YEplac195/sma1 | 1 | Gal-AK2 99 | 2.0 |
2 | YEplac195/sma1 | 1 | GPD-AK2 80 | 2.0 |
3 | YEplac195/sma1 | 1 | Ter-AK2 81 | 2.0 |
4 | YEplac195/sma1 | 1 | T7 AK1 82 | 2.0 |
5 | YEplac195/sma1 | 1 | T7 AK1.2 83 | 2.0 |
6 | YEplac195/sma1 | 1 | Gal1 gfp 85 | 2.0 |
7 | YEplac195/sma1 | 1 | ter gfp 86 | 2.0 |
08/13
Nebuilder
Yoshimoto
Sample | Vectorname | Vector amount (ul) | Insert1 name | Insert1 amount(ul) | Insert2 name | Insert2 amount (ul) | Mastermix amount (ul) | Total amount(ul) |
---|
1 | YEPlac195/sma1 | 0.25 | GPD-AK | 1.5 | Ter-AK | 5.0 | 5.0 | 12.0 |
2 | YEPlac195/sma1 | 0.25 | Gal-AK | 2.0 | Ter-AK | 5.0 | 5.0 | 12.0 |
08/13
Transformation
Yoshimoto
sample | CompetentCell name | DNA name |
---|
1 | DH5a | Nebuilder product |
2 | DH5a | Nebuilder product |
3 | DH5a | Nebuilder product |
4 | DH5a | Nebuilder product |
08/14
PCR
Ohara
lane | Primer Foward | Primer Revese | Template |
---|
1 | M13F | M13R | 79-1 |
2 | M13F | M13R | 80-1 |
3 | M13F | M13R | 80-2 |
4 | M13F | M13R | 81-1 |
5 | M13F | M13R | 97-1 |
6 | M13F | M13R | 98-1 |
7 | M13F | M13R | 98-2 |
8 | M13F | M13R | 98-3 |
9 | M13F | M13R | 92-1 |
10 | M13F | M13R | 99-1 |
11 | M13F | M13R | 100-1 |
12 | M13F | M13R | 101-1 |
13 | M13F | M13R | 101-2 |
14 | M13F | M13R | 101-3 |
15 | M13F | M13R | 102-1 |
16 | M13F | M13R | water |
PCR Polymerase : 2*KAPA hifi HS readymix
95.0°C : 3min
98.0°C : 20sec
60.0°C : 15sec
72.0°C : 1min20sec
72.0°C : 1min
4.0°C : Infinate
X35 cycle
08/14
Restriction Enzyme digestion
Ohara
lane | Sample DNA name | DNA amount (ul) | RE1 name | RE1 amount(ul) | RE2 name | RE2 amount(ul) | Buffer name | Buffer amount(ul) | Total volume(ul) |
---|
1 | 2 | 5.0 | Pst1 | 1.0 | None | None | 10xH | 1.0 | 10.0 |
2 | 3 | 5.0 | Pst1 | 1.0 | None | None | 10xH | 1.0 | 10.0 |
3 | 4 | 5.0 | Pst1 | 1.0 | None | None | 10xH | 1.0 | 10.0 |
4 | 5 | 5.0 | Pst1 | 1.0 | None | None | 10xH | 1.0 | 10.0 |
5 | 7 | 5.0 | Pst1 | 1.0 | None | None | 10xH | 1.0 | 10.0 |
6 | 14 | 5.0 | Pst1 | 1.0 | None | None | 10xH | 1.0 | 10.0 |
7 | 16 | 5.0 | Pst1 | 1.0 | None | None | 10xH | 1.0 | 10.0 |
8 | 22 | 5.0 | Pst1 | 1.0 | None | None | 10xH | 1.0 | 10.0 |
9 | 77-1 | 5.0 | Pst1 | 1.0 | EcoR1 | 1.0 | 10xH | 1.0 | 10.0 |
10 | 77-2 | 5.0 | Pst1 | 1.0 | EcoR1 | 1.0 | 10xH | 1.0 | 10.0 |
11 | 77-3 | 5.0 | Pst1 | 1.0 | EcoR1 | 1.0 | 10xH | 1.0 | 10.0 |
12 | 24 | 5.0 | Pst1 | 1.0 | None | None | 10xH | 1.0 | 10.0 |
13 | 26 | 5.0 | Pst1 | 1.0 | None | None | 10xH | 1.0 | 10.0 |
14 | 30 | 5.0 | Pst1 | 1.0 | None | None | 10xH | 1.0 | 10.0 |
15 | 36 | 5.0 | Pst1 | 1.0 | None | None | 10xH | 1.0 | 10.0 |
16 | 78-1 | 5.0 | Pst1 | 1.0 | None | None | 10xH | 1.0 | 10.0 |
17 | 78-2 | 5.0 | Pst1 | 1.0 | None | None | 10xH | 1.0 | 10.0 |
18 | 78 | 5.0 | Pst1 | 1.0 | None | None | 10xH | 1.0 | 10.0 |
19 | 99 | 5.0 | Pst1 | 1.0 | None | None | 10xH | 1.0 | 10.0 |
20 | 100 | 5.0 | Pst1 | 1.0 | None | None | 10xH | 1.0 | 10.0 |
21 | 101 | 5.0 | Pst1 | 1.0 | None | None | 10xH | 1.0 | 10.0 |
08/14
colony_PCR
Yoshimoto
lane | Primer Foward | Primer Revese | Template |
---|
1 | M13F | M13R | 97 |
2 | M13F | M13R | 98 |
3 | M13F | M13R | 99 |
4 | M13F | M13R | 100 |
5 | M13F | M13R | 101 |
6 | M13F | M13R | 102 |
PCR Polymerase : 2*KAPA hifi HS readymix
95.0°C : 3min
98.0°C : 20sec
60.0°C : 15sec
72.0°C : 1min20sec
72.0°C : 5min
4.0°C : Infinate
X35 cycle
08/14
Transformation
Yoshimoto
sample | CompetentCell name | DNA name |
---|
1 | DH5a | Nebuilder product gal1-gfp |
2 | DH5a | Nebuilder product GPD-gfp |
3 | DH5a | 8/13 ligation product Gal-AK2 |
4 | DH5a | 8/13 ligation product GPD-AK2 |
5 | DH5a | 8/13 ligation product Ter-AK2 |
08/14
Transformation
Yoshimoto
sample | CompetentCell name | DNA name |
---|
1 | DH5a | ligation product Gal-AK2 |
2 | DH5a | ligation product GPD-AK2 |
3 | DH5a | ligation product Ter-AK2 |
08/14
Miniprep_Nanodrop
Yoshimoto
sample | Sample name | 260/280 | 260/230 | ng/ul |
---|
1 | 101 | 1.97 | 0.93 | 24.9 |
2 | 100 | 1.99 | 0.99 | 34.8 |
3 | 97 | 1.94 | 1.35 | 50.1 |
4 | 98 | 1.97 | 1.97 | 40.3 |
5 | 99.0 | 1.96 | 1.65 | 99.7 |
6 | 73.5 | 1.82 | 0.9 | 47.9 |
7 | 78.4 | 1.86 | 0.99 | 43.7 |
8 | 71-3 | 1.88 | 1.21 | 67.4 |
9 | 36 | 1.88 | 1.41 | 129.5 |
10 | 77-1 | 1.81 | 0.99 | 70.8 |
11 | 77-2 | 1.89 | 1.2 | 63.0 |
12 | 30 | 1.82 | 1.23 | 135.8 |
13 | 26 | 1.86 | 1.31 | 143.9 |
14 | 24 | 1.88 | 1.43 | 141.5 |
15 | 14 | 1.82 | 1.31 | 178.7 |
16 | 16 | 1.91 | 1.37 | 132.3 |
17 | 22 | 1.79 | 1.35 | 207.8 |
18 | 7 | 1.85 | 1.28 | 94.4 |
19 | 5 | 1.79 | 0.91 | 63.8 |
20 | 4 | 1.66 | 0.81 | 64.2 |
21 | 2 | 1.87 | 0.96 | 50.5 |
22 | 3 | 1.79 | 1.15 | 67.1 |
The way of extraction : promega
08/15
Restriction Enzyme digestion
Ohara
lane | Sample DNA name | DNA amount (ul) | RE1 name | RE1 amount(ul) | Buffer name | Buffer amount(ul) | Total volume(ul) |
---|
1 | 92 | 5.0 | Pst1 | 1.0 | 10xH | 1.0 | 10.0 |
2 | 97 | 5.0 | Pst1 | 1.0 | 10xH | 1.0 | 10.0 |
3 | 98 | 5.0 | Pst1 | 1.0 | 10xH | 1.0 | 10.0 |
4 | 100 | 5.0 | Pst1 | 1.0 | 10xH | 1.0 | 10.0 |
5 | 101 | 5.0 | Pst1 | 1.0 | 10xH | 1.0 | 10.0 |
6 | 92 | 5.0 | Pst1 | 1.0 | 10xH | 1.0 | 10.0 |
7 | 97 | 5.0 | Pst1 | 1.0 | 10xH | 1.0 | 10.0 |
8 | 98 | 5.0 | Pst1 | 1.0 | 10xH | 1.0 | 10.0 |
9 | 100 | 5.0 | Pst1 | 1.0 | 10xH | 1.0 | 10.0 |
10 | 101 | 5.0 | Pst1 | 1.0 | 10xH | 1.0 | 10.0 |
08/15
Nebuilder
Yoshimoto
Sample | Vectorname | Vector amount (ul) | Insert1 name | Insert1 amount(ul) | Insert2 name | Insert2 amount (ul) | Mastermix amount (ul) | Total amount(ul) |
---|
1 | YEplac195/sma1 | 0.25 | Gal-AK | 1.825 | Ter-AK | 1.5 | 5.0 | 10.0 |
2 | YEplac195/sma1 | 0.25 | GPD-AK | 1.25 | Ter-AK | 1.5 | 5.0 | 10.0 |
08/15
Transformation
Yoshimoto
sample | CompetentCell name | DNA name |
---|
1 | DH5a | 111 |
2 | DH5a | 112 |
08/17
PCR
Yoshimoto
lane | Primer Foward | Primer Revese | Template |
---|
1 | IK25 | IK26 | Rev |
2 | IK27 | IK22 | yeast genome |
3 | IK23 | IK28 | yeast genome |
PCR Polymerase : 2*KAPA hifi HS Ready mix
98.0°C : 3min
98.0°C : 20sec
62.0°C : 30sec
72.0°C : 1min
72.0°C : 5min
4.0°C : Infinate
X35 cycle
08/17
PCR
Yoshimoto
lane | Primer Foward | Primer Revese | Template |
---|
1 | IK27 | IK22 | yeast genome |
2 | IK23 | IK28 | yeast genome |
3 | IK27 | IK28 | GPDp+Rev+GPDt |
PCR Polymerase : 2*KAPA hifi HS ready mix
98.0°C : 3min
98.0°C : 20sec
60.0°C : 30sec
72.0°C : 1min
72.0°C : 5min
4.0°C : Infinate
X35 cycle
08/17
PCR
Ohara
lane | Primer Foward | Primer Revese | Template |
---|
1 | IK25 | IK26 | Rev |
2 | IK27 | IK22 | yeast genome |
3 | IK23 | IK28 | yeast genome |
4 | IK25 | IK26 | Rev after purification |
5 | IK27 | IK22 | GPDp after purification |
6 | IK23 | IK28 | GPDt after purification |
PCR Polymerase : 2*KAPA hifi HS ready mix
95.0°C : 3min
98.0°C : 20sec
60.0°C : 15sec
72.0°C : 40sec
72.0°C : 5min
4.0°C : Infinate
X36 cycle
08/17
PCR
Yoshimoto
lane | Primer Foward | Primer Revese | Template |
---|
1 | M13F | M13R | 86-1 |
2 | M13F | M13R | 86-2 |
3 | M13F | M13R | 81-1 |
4 | M13F | M13R | 81-2 |
5 | M13F | M13R | 81-3 |
6 | M13F | M13R | 95-1 |
7 | M13F | M13R | 95-2 |
8 | M13F | M13R | 95-3 |
9 | M13F | M13R | 95-4 |
10 | M13F | M13R | 93-1 |
11 | M13F | M13R | 93-2 |
12 | M13F | M13R | 92 |
13 | M13F | M13R | brue colony |
14 | M13F | M13R | 11 |
PCR Polymerase : 2*KAPA hifi HS readymix
95.0°C : 3min
95.0°C : 20sec
60.0°C : 15sec
72.0°C : 1min20sec
72.0°C : 1min
4.0°C : Infinate
X35 cycle
08/17
Nebuilder
Yoshimoto
Sample | Vectorname | Vector amount (ul) | Insert1 name | Insert1 amount(ul) | Insert2 name | Insert2 amount (ul) | Mastermix amount (ul) | Total amount(ul) |
---|
1 | YEplac195/sma1 | 0.8 | Gal-AK2 | 6.5 | Ter-AK2 | 5.0 | 10.0 | 22.3 |
2 | YEplac195/sma1 | 0.8 | GPD-AK2 | 4.2 | Ter-AK2 | 5.0 | 10.0 | 20.0 |
08/17
Transformation
Yoshimoto
sample | CompetentCell name | DNA name |
---|
1 | BL21 | 113 |
2 | BL21 | 114 |
3 | BL21 | 115 |
08/18
Restriction Enzyme digestion
Ohara
lane | Sample DNA name | DNA amount (ul) | RE1 name | RE1 amount(ul) | RE2 name | RE2 amount(ul) | Buffer name | Buffer amount(ul) | Total volume(ul) |
---|
1 | 79-2 | 5.0 | Pst1 | 1.0 | EcoR1 | 1.0 | 10xH | 1.0 | 10.0 |
2 | 79-3 | 5.0 | Pst1 | 1.0 | EcoR1 | 1.0 | 10xH | 1.0 | 10.0 |
3 | 79-6 | 5.0 | Pst1 | 1.0 | EcoR1 | 1.0 | 10xH | 1.0 | 10.0 |
4 | 80-2 | 5.0 | Pst1 | 1.0 | EcoR1 | 1.0 | 10xH | 1.0 | 10.0 |
5 | 80-7 | 5.0 | Pst1 | 1.0 | EcoR1 | 1.0 | 10xH | 1.0 | 10.0 |
6 | 80-13 | 5.0 | Pst1 | 1.0 | EcoR1 | 1.0 | 10xH | 1.0 | 10.0 |
7 | 81-1 | 5.0 | Pst1 | 1.0 | None | None | 10xH | 1.0 | 10.0 |
8 | 81-2 | 5.0 | Pst1 | 1.0 | None | None | 10xH | 1.0 | 10.0 |
9 | 97 | 5.0 | Pst1 | 1.0 | None | None | 10xH | 1.0 | 10.0 |
10 | 99-A | 5.0 | Pst1 | 1.0 | None | None | 10xH | 1.0 | 10.0 |
11 | 99-B | 5.0 | Pst1 | 1.0 | None | None | 10xH | 1.0 | 10.0 |
12 | 100 | 5.0 | Pst1 | 1.0 | None | None | 10xH | 1.0 | 10.0 |
13 | 100-1 | 5.0 | Pst1 | 1.0 | None | None | 10xH | 1.0 | 10.0 |
14 | 100-2 | 5.0 | Pst1 | 1.0 | None | None | 10xH | 1.0 | 10.0 |
15 | 101-1 | 5.0 | Pst1 | 1.0 | None | None | 10xH | 1.0 | 10.0 |
16 | 101 3 | 5.0 | Pst1 | 1.0 | None | None | 10xH | 1.0 | 10.0 |
17 | 102 | 5.0 | Pst1 | 1.0 | None | None | 10xH | 1.0 | 10.0 |
08/18
PCR
Yoshimoto
lane | Primer Foward | Primer Revese | Template |
---|
1 | IK27 | IK28 | Rev |
PCR Polymerase : 2*KAPA hifi HS Ready mix
98.0°C : 3min
98.0°C : 20sec
60.0°C : 20sec
72.0°C : 1min20sec
72.0°C : 5min
4.0°C : Infinate
X35 cycle
08/18
PCR
Ohara
lane | Primer Foward | Primer Revese | Template |
---|
1 | M13F | AK2R | 79-2 |
2 | M13F | AK2R | 79-3 |
3 | M13F | AK2R | 80-13 |
4 | M13F | AK2F | 79-2 |
5 | M13F | AK2F | 79-3 |
6 | M13F | AK2F | 80-13 |
7 | M13R | M13F | blue colony |
PCR Polymerase : 2*KAPA hifi HS readymix
95.0°C : 3min
98.0°C : 20sec
60.0°C : 15sec
72.0°C : 1min30sec
72.0°C : 5min
4.0°C : Infinate
X35 cycle
08/18
PCR
Ohara
lane | Primer Foward | Primer Revese | Template |
---|
1 | IK22 | IK27 | GPDp after purification |
2 | IK22 | IK27 | GPDp after purification |
3 | IK22 | IK27 | GPDp after purification |
4 | IK22 | IK27 | GPDp after purification |
5 | M13F | AK2R | 113gal |
6 | M13F | AK2R | 114GPD |
PCR Polymerase : 2*KAPA hifi HS readymix
95.0°C : 3min
98.0°C : 20sec
60.0°C : 15sec
72.0°C : 1min30sec
72.0°C : 5min
4.0°C : Infinate
X35 cycle
PCR pulification: lane1~4
Gel extraction: lane1~4
08/19
Ligation
Yoshimoto
sample | Vector name | Vector amount(ul) | Insert name | Insert amount(ul) |
---|
1 | 141 | 0.34 | 2T7akRRE3' | 9.4 |
2 | 142 | 0.34 | 2T7ak1stem | 8.3 |
3 | 143 | 0.34 | 1T7ak1stem | 4.4 |
4 | 144 | 0.34 | 2T7RREak1 | 5.3 |
5 | 145 | 0.34 | 1T7RREak1 | 8.5 |
6 | 146 | 0.34 | GPD+AK2 | 13.2 |
7 | 147 | 0.34 | Gal1+AK2 | 15.8 |
8 | 148 | 0.34 | AK2+ter | 20.1 |
08/20
PCR
Ohara
lane | Primer Foward | Primer Revese | Template |
---|
1 | IK22 | IK27 | GPDp |
2 | IK22 | IK27 | GPDp |
3 | IK22 | IK27 | yeast genome |
4 | IK22 | IK27 | yeast genome |
5 | IK22 | IK27 | yeast genome |
6 | IK22 | IK27 | yeast genome |
7 | IK27 | IK28 | GPDp+Rev+GPDt |
8 | IK27 | IK28 | GPDp+Rev+GPDt |
PCR Polymerase : 2*KAPA hifi HS ready mix
95.0°C : 3min
98.0°C : 20sec
60.0°C : 15sec
72.0°C : 1min30sec
72.0°C : 5min
4.0°C : Infinate
X35 cycle
08/20
PCR
Yoshimoto
lane | Primer Foward | Primer Revese | Template |
---|
1 | IK27 | IK22 | GPDp |
2 | IK21 | IK22 | GPDp |
PCR Polymerase : 2*KAPA HS readymix
98.0°C : 3min
98.0°C : 20sec
60.0°C : 30sec
72.0°C : 1min
72.0°C : 5min
4.0°C : Infinate
X35 cycle
08/21
PCR
Ohara
lane | Primer Foward | Primer Revese | Template |
---|
1 | IK29 | IK30 | Rev |
2 | IK29 | IK30 | Rev |
3 | IK29 | IK30 | Rev |
4 | IK29 | IK30 | Rev |
5 | M13R | AK1R | 141 1-1 |
6 | M13R | AK1R | 141 1-2 |
7 | M13R | AK1R | 141 1-3 |
8 | M13R | AK1R | 141 1-4 |
9 | M13R | AK1R | 142 1-1 |
10 | M13R | AK1R | 142 1-2 |
11 | M13R | AK1R | 142 1-3 |
12 | M13R | AK1R | 142 1-4 |
13 | M13R | AK1F | 141 1-1 |
14 | M13R | AK1F | 141 1-2 |
15 | M13R | AK1F | 141 1-3 |
16 | M13R | AK1F | 141 1-4 |
17 | M13R | AK1F | 142 1-1 |
18 | M13R | AK1F | 142 1-2 |
19 | M13R | AK1F | 142 1-3 |
20 | M13R | AK1F | 142 1-4 |
PCR Polymerase : 2*KAPA hifi HS Readymix
95.0°C : 3min
98.0°C : 20sec
60.0°C : 15sec
72.0°C : 1min20sec
72.0°C : 5min
4.0°C : Infinate
X35 cycle
08/21
PCR
Ohara
lane | Primer Foward | Primer Revese | Template |
---|
1 | M13F | AK1R | 141 2-1 |
2 | M13F | AK1R | 141 2-4 |
3 | M13F | AK1R | 141 1-15 |
4 | M13F | AK1R | 142 1-5 |
5 | M13F | AK1R | 143 9 |
6 | M13F | AK1R | 143 13 |
7 | M13F | AK1R | 143 27 |
8 | M13F | AK1R | 143 2 |
9 | M13F | AK1F | 141 2-1 |
10 | M13F | AK1F | 141 2-4 |
11 | M13F | AK1F | 141 1-15 |
12 | M13F | AK1F | 142 1-5 |
13 | M13F | AK1F | 143 9 |
14 | M13F | AK1F | 143 13 |
15 | M13F | AK1F | 143 27 |
16 | M13F | AK1F | 143 2 |
PCR Polymerase : 2*KAPA hifi HS Readymix
95.0°C : 3min
98.0°C : 20sec
60.0°C : 15sec
72.0°C : 1min20sec
72.0°C : 5min
4.0°C : Infinate
X35 cycle
08/21
PCR
Ohara
lane | Primer Foward | Primer Revese | Template |
---|
1 | IK29 | IK30 | Rev |
2 | IK29 | IK30 | Rev |
3 | IK29 | IK30 | Rev |
4 | IK29 | IK30 | Rev |
PCR Polymerase : 2*KAPA hifi HS Readymix
95.0°C : 3min
98.0°C : 20sec
60.0°C : 15sec
72.0°C : 1min20sec
72.0°C : 5min
4.0°C : Infinate
X35 cycle
PCR pulification: lane2~4
Gel extraction: lane2~4
08/22
PCR
Ohara
lane | Primer Foward | Primer Revese | Template |
---|
1 | IK21 | IK22 | GPDp |
2 | IK21 | IK22 | GPDp |
3 | IK21 | IK22 | GPDp |
4 | IK21 | IK22 | GPDp |
5 | IK21 | IK22 | water |
6 | IK23 | IK28 | GPDt |
7 | IK23 | IK28 | GPDt |
8 | IK23 | IK28 | GPDt |
9 | IK23 | IK28 | GPDt |
10 | IK23 | IK28 | water |
PCR Polymerase : 2*KAPA hifi HS Readymix
98.0°C : 3min
98.0°C : 20sec
60.0°C : 30sec
72.0°C : 1min
72.0°C : 5min
4.0°C : Infinate
X35 cycle
PCR pulification: lane2~4
Gel extraction: lane2~4
08/22
PCR
Ohara
lane | Primer Foward | Primer Revese | Template |
---|
1 | IK27 | IK28 | GPDp+Rev+GPDt |
2 | IK27 | IK28 | GPDp+Rev+GPDt |
3 | IK23 | IK24 | yeast genome |
4 | IK23 | IK24 | yeast genome |
5 | IK23 | IK24 | yeast genome |
6 | IK23 | IK24 | yeast genome |
7 | IK21 | IK24 | GPDp+Rev+GPDt |
PCR Polymerase : 2*KAPA hifi HS ReadyMIx
95.0°C : 3min
98.0°C : 20sec
60.0°C : 15sec
72.0°C : 1min30sec
72.0°C : 5min
4.0°C : Infinate
X35 cycle
08/22
PCR
Fukuda
lane | Primer Foward | Primer Revese | Template |
---|
1 | M13F | M13R | 144 11 |
2 | M13F | M13R | 144 3 |
3 | M13F | M13R | 144 7 |
4 | M13F | M13R | 144 1 |
5 | M13F | M13R | 144 17 |
6 | M13F | M13R | 144 5 |
7 | M13F | M13R | 144 9 |
8 | M13F | M13R | 144 13 |
9 | M13F | M13R | 142 1 |
10 | M13F | M13R | 142 5 |
11 | M13F | M13R | 142 3 |
12 | M13F | M13R | 142 4 |
13 | M13F | M13R | 142 2-1 |
14 | M13F | M13R | 142 2-2 |
15 | M13F | M13R | 142 2-3 |
16 | M13F | M13R | 142 2-4 |
17 | M13F | M13R | 143 13 |
18 | M13F | M13R | 143 15 |
19 | M13F | M13R | 143 5 |
20 | M13F | M13R | 153 21 |
21 | M13F | M13R | 143 17 |
22 | M13F | M13R | 143 19 |
23 | M13F | M13R | 143 25 |
24 | M13F | M13R | 143 9 |
PCR Polymerase : 2*KAPA hifi HS ReadyMix
95.0°C : 3min
98.0°C : 20sec
60.0°C : 15sec
72.0°C : 1min30sec
72.0°C : 5min
4.0°C : Infinate
X35 cycle
08/22
PCR
Takimoto
lane | Primer Foward | Primer Revese | Template |
---|
1 | M13F | M13R | 145 1-13 |
2 | M13F | M13R | 145 2-13 |
3 | M13F | M13R | 145 2-15 |
4 | M13F | M13R | 145 1-5 |
5 | M13F | M13R | 145 2-17 |
6 | M13F | M13R | 145 2-11 |
7 | M13F | M13R | 145 2-9 |
8 | M13F | M13R | 145 1-17 |
9 | M13F | M13R | 146 2-3 |
10 | M13F | M13R | 146 2-1 |
11 | M13F | M13R | 146 1-3 |
12 | M13F | M13R | 146 2-7 |
13 | M13F | M13R | 146 2-9 |
14 | M13F | M13R | 146 2-5 |
15 | M13F | M13R | 146 1-4 |
PCR Polymerase : 2*KAPA hifi HS ReadyMix
95.0°C : 3min
98.0°C : 20sec
60.0°C : 15sec
72.0°C : 1min30sec
72.0°C : 5min
4.0°C : Infinate
X35 cycle
08/22
Nebuilder
Yoshimoto
Sample | Vectorname | Vector amount (ul) | Insert1 name | Insert1 amount(ul) | Insert2 name | Insert2 amount (ul) | Mastermix amount (ul) | Total amount(ul) |
---|
1 | 171.0 | Vector amount (ul) | Gal1+gfp-2 | 0.78 | Gal1+gfp | 1.03 | 10.0 | Total amount(ul) |
2 | 172.0 | Vector amount (ul) | Gal1+AK2 | 1.26 | Gal1+AK2-2 | 1.01 | 10.0 | Total amount(ul) |
3 | 173.0 | Vector amount (ul) | 164.0 | 0.65 | 165.0 | 0.57 | 10.0 | Total amount(ul) |
4 | 174.0 | Vector amount (ul) | 161.0 | 0.65 | 162.0 | 0.76 | 10.0 | Total amount(ul) |
5 | 175.0 | Vector amount (ul) | 161.0 | 0.38 | 167.0 | 0.83 | 10.0 | Total amount(ul) |
6 | 176.0 | Vector amount (ul) | 166.0 | 0.38 | 165.0 | 0.83 | 10.0 | Total amount(ul) |
7 | 177.0 | Vector amount (ul) | 163.0 | 0.38 | 162.0 | 0.83 | 10.0 | Total amount(ul) |
8 | 178.0 | Vector amount (ul) | 163.0 | 0.38 | 167.0 | 0.83 | 10.0 | Total amount(ul) |
08/22
PCR
Ohara
lane | Primer Foward | Primer Revese | Template |
---|
1 | None | None | GPDp+Rev+GPDt |
2 | IK27 | IK28 | lane1 after 15 cycle |
PCR Polymerase : 2*KAPA hifi Hs ReadyMix
95.0°C : 3min
98.0°C : 20sec
60.0°C : 15sec
72.0°C : 1min30sec
72.0°C : 5min
4.0°C : Infinate
X35 cycle
08/22
PCR
Ohara
lane | Primer Foward | Primer Revese | Template |
---|
1 | IK23 | IK28 | GPDt |
2 | IK23 | IK28 | GPDt |
3 | IK23 | IK28 | GPDt |
4 | IK23 | IK28 | GPDt |
5 | IK27 | IK30 | GPDp + Rev |
6 | IK21 | IK30 | GPDp + Rev |
PCR Polymerase : 2*KAPA hifi Robust
95.0°C : 3min
98.0°C : 20sec
60.0°C : 15sec
72.0°C : 40sec
72.0°C : 5min
4.0°C : Infinate
X35 cycle
08/24
PCR
Yoshimoto
lane | Primer Foward | Primer Revese | Template |
---|
1 | M13F | M13R | 141 2-1 |
2 | M13F | M13R | 141 2-2 |
3 | M13F | M13R | 141 2-4 |
4 | M13F | M13R | 141 2-6 |
5 | M13F | M13R | 142 1-5 |
6 | M13F | M13R | 142 2-4 |
7 | M13F | M13R | 142 1-3 |
8 | M13F | M13R | 142 1-4 |
9 | M13F | M13R | 143 13 |
10 | M13F | M13R | 143 19 |
11 | M13F | M13R | 143 25 |
12 | M13F | M13R | 144 18 |
13 | M13F | M13R | 144 17 |
14 | M13F | M13R | 144 5 |
15 | M13F | M13R | 144 9 |
16 | M13F | M13R | 143 19 |
17 | M13F | M13R | 145 1-5 |
18 | M13F | M13R | 145 2-15 |
19 | M13F | M13R | 143 2-13 |
20 | M13F | M13R | 145 1-13 |
21 | M13F | AK1R | 141 2-1 |
22 | M13F | AK1R | 141 2-2 |
23 | M13F | AK1R | 141 2-4 |
24 | M13F | AK1R | 141 2-6 |
25 | M13F | AK1R | 142 1-5 |
26 | M13F | AK1R | 142 2-4 |
27 | M13F | AK1R | 142 1-3 |
28 | M13F | AK1R | 142 1-4 |
29 | M13F | AK1R | 145 1-5 |
30 | M13F | AK1R | 145 2-15 |
31 | M13F | AK1R | 143 2-13 |
32 | M13F | AK1R | 145 1-13 |
33 | M13F | AK1R | 144 17 |
34 | M13F | AK1R | 144 5 |
35 | M13F | AK1R | 144 9 |
36 | M13F | AK1R | 143 19 |
37 | M13F | AK1R | 143 13 |
38 | M13F | AK1R | 143 19 |
39 | M13F | AK1R | 143 25 |
40 | M13F | AK1R | 144 9 |
PCR Polymerase : 2*KAPA Robust
98.0°C : 3min
98.0°C : 20sec
60.0°C : 30sec
72.0°C : 40sec
72.0°C : 5min
4.0°C : Infinate
X35 cycle
08/26
PCR
Ohara
lane | Primer Foward | Primer Revese | Template |
---|
1 | IK27 | IK30 | GPDp + Rev after gel extraction |
2 | IK27 | IK30 | GPDp + Rev after gel extraction |
3 | IK27 | IK30 | GPDp + Rev after gel extraction |
4 | IK27 | IK30 | GPDp + Rev after gel extraction |
5 | IK23 | IK28 | GPDt after PCR purification |
6 | IK23 | IK28 | GPDt after PCR purification |
7 | IK23 | IK28 | GPDt after PCR purification |
8 | IK23 | IK28 | GPDt after PCR purification |
PCR Polymerase : 2*KAPA hifi HS Readymix
98.0°C : 3min
98.0°C : 20sec
60.0°C : 30sec
72.0°C : 40sec
72.0°C : 5min
4.0°C : Infinate
X35 cycle
08/26
PCR
Yoshimoto
lane | Primer Foward | Primer Revese | Template |
---|
1 | M13F | M13R | 145-2 11 |
2 | M13F | M13R | 145-1 9 |
3 | M13F | M13R | 145 2-7 |
4 | M13F | M13R | 145 1-21 |
5 | M13F | M13R | 145 1-3 |
6 | M13F | M13R | 145 2-3 |
7 | M13F | M13R | 145 1-19 |
8 | M13F | M13R | 142 1-2 |
9 | M13F | AK1F | 145-2 11 |
10 | M13F | AK1F | 145-1 9 |
11 | M13F | AK1F | 145 2-7 |
12 | M13F | AK1F | 145 1-21 |
13 | M13F | AK1F | 145 1-3 |
14 | M13F | AK1F | 145 2-3 |
15 | M13F | AK1F | 145 1-19 |
16 | M13F | AK1F | 142 1-2 |
PCR Polymerase : 2*KAPA hifi
98.0°C : 3min
98.0°C : 20sec
60.0°C : 15sec
72.0°C : 25sec
72.0°C : 3min
4.0°C : Infinate
X35 cycle
08/26
Restriction Enzyme digestion
Ohara
lane | Sample DNA name | DNA amount (ul) | RE1 name | RE1 amount(ul) | Buffer name | Buffer amount(ul) | Total volume(ul) |
---|
1 | 114-1 | 2.0 | Pst1 | 1.0 | 10xH | 1.0 | 10.0 |
2 | 114-2 | 2.0 | Pst1 | 1.0 | 10xH | 1.0 | 10.0 |
3 | 172 | 2.0 | Pst1 | 1.0 | 10xH | 1.0 | 10.0 |
4 | 172-1 | 2.0 | Pst1 | 1.0 | 10xH | 1.0 | 10.0 |
5 | 172-2 | 2.0 | Pst1 | 1.0 | 10xH | 1.0 | 10.0 |
6 | 172-4 | 2.0 | Pst1 | 1.0 | 10xH | 1.0 | 10.0 |
7 | 175-3 | 2.0 | Pst1 | 1.0 | 10xH | 1.0 | 10.0 |
8 | 175-10 | 2.0 | Pst1 | 1.0 | 10xH | 1.0 | 10.0 |
9 | 176 | 2.0 | Pst1 | 1.0 | 10xH | 1.0 | 10.0 |
10 | 177 | 2.0 | Pst1 | 1.0 | 10xH | 1.0 | 10.0 |
08/27
Nebuilder
Yoshimoto
Sample | Vectorname | Vector amount (ul) | Insert1 name | Insert1 amount(ul) | Insert2 name | Insert2 amount (ul) | Mastermix amount (ul) | Total amount(ul) |
---|
1 | Rev | 3.02 | GPDp+Rev | 0.669 | GPDt | 0.875 | 10.0 | Total amount(ul) |
2 | 173.0 | 3.02 | 164.0 | 0.325 | 165.0 | 0.285 | 5.0 | Total amount(ul) |
3 | 175.0 | 3.02 | 161.0 | 0.19 | 167.0 | 0.42 | 5.0 | Total amount(ul) |
4 | 178.0 | 3.02 | 163.0 | 0.41 | 167.0 | 0.42 | 5.0 | Total amount(ul) |
08/27
Restriction Enzyme digestion
Ohara
lane | Sample DNA name | DNA amount (ul) | RE1 name | RE1 amount(ul) | Buffer name | Buffer amount(ul) | Total volume(ul) |
---|
1 | 171-1 | 5.0 | Pst1 | 1.0 | 10xH | 1.0 | 10.0 |
2 | 171-1-2 | 5.0 | Pst1 | 1.0 | 10xH | 1.0 | 10.0 |
3 | 172-2 | 5.0 | Pst1 | 1.0 | 10xH | 1.0 | 10.0 |
4 | 172-2-2 | 5.0 | Pst1 | 1.0 | 10xH | 1.0 | 10.0 |
5 | 172-3 | 5.0 | Pst1 | 1.0 | 10xH | 1.0 | 10.0 |
6 | 172-4 | 5.0 | Pst1 | 1.0 | 10xH | 1.0 | 10.0 |
7 | 172-5 | 5.0 | Pst1 | 1.0 | 10xH | 1.0 | 10.0 |
8 | 172-6 | 5.0 | Pst1 | 1.0 | 10xH | 1.0 | 10.0 |
9 | 172-8 | 5.0 | Pst1 | 1.0 | 10xH | 1.0 | 10.0 |
10 | 172-9 | 5.0 | Pst1 | 1.0 | 10xH | 1.0 | 10.0 |
11 | 172-10 | 5.0 | Pst1 | 1.0 | 10xH | 1.0 | 10.0 |
12 | 172-11 | 5.0 | Pst1 | 1.0 | 10xH | 1.0 | 10.0 |
13 | 172-12 | 5.0 | Pst1 | 1.0 | 10xH | 1.0 | 10.0 |
14 | 172-13 | 5.0 | Pst1 | 1.0 | 10xH | 1.0 | 10.0 |
15 | 175-1 | 5.0 | EcoR1 | 1.0 | 10xH | 1.0 | 10.0 |
16 | 175-2 | 5.0 | EcoR1 | 1.0 | 10xH | 1.0 | 10.0 |
17 | 175-3 | 5.0 | EcoR1 | 1.0 | 10xH | 1.0 | 10.0 |
18 | 175-4 | 5.0 | EcoR1 | 1.0 | 10xH | 1.0 | 10.0 |
19 | 176-1 | 5.0 | EcoR1 | 1.0 | 10xH | 1.0 | 10.0 |
20 | 177-2 | 5.0 | EcoR1 | 1.0 | 10xH | 1.0 | 10.0 |
21 | 177-3 | 5.0 | EcoR1 | 1.0 | 10xH | 1.0 | 10.0 |
22 | 177-4 | 5.0 | EcoR1 | 1.0 | 10xH | 1.0 | 10.0 |
23 | 177-5 | 5.0 | EcoR1 | 1.0 | 10xH | 1.0 | 10.0 |
24 | 177-1 | 5.0 | EcoR1 | 1.0 | 10xH | 1.0 | 10.0 |
25 | 177-1-2 | 5.0 | EcoR1 | 1.0 | 10xH | 1.0 | 10.0 |
26 | 171-1 | 5.0 | EcoR1 | 1.0 | 10xH | 1.0 | 10.0 |
27 | 172-2 | 5.0 | EcoR1 | 1.0 | 10xH | 1.0 | 10.0 |
28 | 175-1 | 5.0 | EcoR1 | 1.0 | 10xH | 1.0 | 10.0 |
29 | 175-2 | 5.0 | EcoR1 | 1.0 | 10xH | 1.0 | 10.0 |
30 | 175-3 | 5.0 | EcoR1 | 1.0 | 10xH | 1.0 | 10.0 |
31 | 175-4 | 5.0 | EcoR1 | 1.0 | 10xH | 1.0 | 10.0 |
32 | 176-1 | 5.0 | EcoR1 | 1.0 | 10xH | 1.0 | 10.0 |
33 | 176-2 | 5.0 | EcoR1 | 1.0 | 10xH | 1.0 | 10.0 |
34 | 177-2 | 5.0 | EcoR1 | 1.0 | 10xH | 1.0 | 10.0 |
35 | 177-3 | 5.0 | EcoR1 | 1.0 | 10xH | 1.0 | 10.0 |
36 | 177-4 | 5.0 | EcoR1 | 1.0 | 10xH | 1.0 | 10.0 |
37 | 177-5 | 5.0 | EcoR1 | 1.0 | 10xH | 1.0 | 10.0 |
08/27
Restriction Enzyme digestion
Ohara
lane | Sample DNA name | DNA amount (ul) | RE1 name | RE1 amount(ul) | Buffer name | Buffer amount(ul) | Total volume(ul) |
---|
1 | 171 1-1 | 5.0 | Pst1 | 1.0 | 10xH | 1.0 | 10.0 |
2 | 171 1-2 | 5.0 | Pst1 | 1.0 | 10xH | 1.0 | 10.0 |
3 | 171 2 | 5.0 | Pst1 | 1.0 | 10xH | 1.0 | 10.0 |
4 | 171 3 | 5.0 | Pst1 | 1.0 | 10xH | 1.0 | 10.0 |
5 | 172-1 | 5.0 | Pst1 | 1.0 | 10xH | 1.0 | 10.0 |
6 | 172-2 | 5.0 | Pst1 | 1.0 | 10xH | 1.0 | 10.0 |
7 | 172-3 | 5.0 | Pst1 | 1.0 | 10xH | 1.0 | 10.0 |
8 | 172-4-1 | 5.0 | Pst1 | 1.0 | 10xH | 1.0 | 10.0 |
9 | 172-4-2 | 5.0 | Pst1 | 1.0 | 10xH | 1.0 | 10.0 |
10 | 172-5 | 5.0 | Pst1 | 1.0 | 10xH | 1.0 | 10.0 |
11 | 172-6 | 5.0 | Pst1 | 1.0 | 10xH | 1.0 | 10.0 |
12 | 172-8 | 5.0 | Pst1 | 1.0 | 10xH | 1.0 | 10.0 |
13 | 172-9 | 5.0 | Pst1 | 1.0 | 10xH | 1.0 | 10.0 |
14 | 172-10 | 5.0 | Pst1 | 1.0 | 10xH | 1.0 | 10.0 |
15 | 172-11 | 5.0 | Pst1 | 1.0 | 10xH | 1.0 | 10.0 |
16 | 172-12 | 5.0 | Pst1 | 1.0 | 10xH | 1.0 | 10.0 |
17 | 172-13 | 5.0 | Pst1 | 1.0 | 10xH | 1.0 | 10.0 |
18 | 171-1-1 | 5.0 | EcoR1 | 1.0 | 10xH | 1.0 | 10.0 |
19 | 174-1 | 5.0 | EcoR1 | 1.0 | 10xH | 1.0 | 10.0 |
20 | 175-1 | 5.0 | EcoR1 | 1.0 | 10xH | 1.0 | 10.0 |
21 | 175-2 | 5.0 | EcoR1 | 1.0 | 10xH | 1.0 | 10.0 |
22 | 175-3 | 5.0 | EcoR1 | 1.0 | 10xH | 1.0 | 10.0 |
23 | 175-4 | 5.0 | EcoR1 | 1.0 | 10xH | 1.0 | 10.0 |
24 | 176-1 | 5.0 | EcoR1 | 1.0 | 10xH | 1.0 | 10.0 |
25 | 176-2 | 5.0 | EcoR1 | 1.0 | 10xH | 1.0 | 10.0 |
26 | 177-2 | 5.0 | EcoR1 | 1.0 | 10xH | 1.0 | 10.0 |
27 | 177-3 | 5.0 | EcoR1 | 1.0 | 10xH | 1.0 | 10.0 |
28 | 177-4 | 5.0 | EcoR1 | 1.0 | 10xH | 1.0 | 10.0 |
29 | 177-5 | 5.0 | EcoR1 | 1.0 | 10xH | 1.0 | 10.0 |
08/28
Restriction Enzyme digestion
Yoshimoto
lane | Sample DNA name | DNA amount (ul) | RE1 name | RE1 amount(ul) | RE2 name | RE2 amount(ul) | Buffer name | Buffer amount(ul) | Total volume(ul) |
---|
1 | 145-2 | 5.0 | Xba1 | 0.5 | EcoR1 | 0.5 | 10xM | 1.0 | 10.0 |
2 | 142 2-11 | 5.0 | Xba1 | 0.5 | EcoR1 | 0.5 | 10xM | 1.0 | 10.0 |
3 | 142 2-2 | 5.0 | Xba1 | 0.5 | EcoR1 | 0.5 | 10xM | 1.0 | 10.0 |
4 | 142 2-1 | 5.0 | Xba1 | 0.5 | EcoR1 | 0.5 | 10xM | 1.0 | 10.0 |
5 | 142 2-3 | 5.0 | Xba1 | 0.5 | EcoR1 | 0.5 | 10xM | 1.0 | 10.0 |
6 | 242 1-4 | 5.0 | Xba1 | 0.5 | EcoR1 | 0.5 | 10xM | 1.0 | 10.0 |
7 | 142 2-4 | 5.0 | Xba1 | 0.5 | EcoR1 | 0.5 | 10xM | 1.0 | 10.0 |
8 | 142 1-5 | 5.0 | Xba1 | 0.5 | EcoR1 | 0.5 | 10xM | 1.0 | 10.0 |
9 | 142 1-3 | 5.0 | Xba1 | 0.5 | EcoR1 | 0.5 | 10xM | 1.0 | 10.0 |
10 | 142 2-5 | 5.0 | Xba1 | 0.5 | EcoR1 | 0.5 | 10xM | 1.0 | 10.0 |
11 | 142 1-2 | 5.0 | Xba1 | 0.5 | EcoR1 | 0.5 | 10xM | 1.0 | 10.0 |
12 | 142 1-1 | 5.0 | Xba1 | 0.5 | EcoR1 | 0.5 | 10xM | 1.0 | 10.0 |
13 | 145 2-3 | 5.0 | Xba1 | 0.5 | EcoR1 | 0.5 | 10xM | 1.0 | 10.0 |
14 | 145 1-2 | 5.0 | Xba1 | 0.5 | EcoR1 | 0.5 | 10xM | 1.0 | 10.0 |
15 | 145 2-7 | 5.0 | Xba1 | 0.5 | EcoR1 | 0.5 | 10xM | 1.0 | 10.0 |
16 | 145 4-9 | 5.0 | Xba1 | 0.5 | EcoR1 | 0.5 | 10xM | 1.0 | 10.0 |
17 | 145 1-11 | 5.0 | Xba1 | 0.5 | EcoR1 | 0.5 | 10xM | 1.0 | 10.0 |
18 | 145 1-3 | 5.0 | Xba1 | 0.5 | EcoR1 | 0.5 | 10xM | 1.0 | 10.0 |
19 | 145 2-11 | 5.0 | Xba1 | 0.5 | EcoR1 | 0.5 | 10xM | 1.0 | 10.0 |
20 | 145 2-5 | 5.0 | Xba1 | 0.5 | EcoR1 | 0.5 | 10xM | 1.0 | 10.0 |
21 | 145 2-8 | 5.0 | Xba1 | 0.5 | EcoR1 | 0.5 | 10xM | 1.0 | 10.0 |
22 | 145 1-14 | 5.0 | Xba1 | 0.5 | EcoR1 | 0.5 | 10xM | 1.0 | 10.0 |
23 | 145 2-15 | 5.0 | Xba1 | 0.5 | EcoR1 | 0.5 | 10xM | 1.0 | 10.0 |
24 | 145 2- 4-2 | 5.0 | Xba1 | 0.5 | EcoR1 | 0.5 | 10xM | 1.0 | 10.0 |
25 | 145 1-17 | 5.0 | Xba1 | 0.5 | EcoR1 | 0.5 | 10xM | 1.0 | 10.0 |
26 | 145 2-17 | 5.0 | Xba1 | 0.5 | EcoR1 | 0.5 | 10xM | 1.0 | 10.0 |
08/29
Restriction Enzyme digestion
Yoshimoto
lane | Sample DNA name | DNA amount (ul) | RE1 name | RE1 amount(ul) | RE2 name | RE2 amount(ul) | Buffer name | Buffer amount(ul) | Total volume(ul) |
---|
1 | 142 1-3 | 5.0 | EcoR1 | 0.5 | Not1 | 0.5 | 10xH, BSA | 1ul 1ul | 10.0 |
2 | 145 1-11 | 5.0 | Xba1 | 0.5 | Not1 | 0.5 | 10xK, BSA | 1ul 1ul | 9.0 |
3 | 145 1-3 | 5.0 | Xba1 | 0.5 | Not1 | 0.5 | 10xK, BSA | 1ul 1ul | 9.0 |
4 | 145 2-42 | 5.0 | Xba1 | 0.5 | Not1 | 0.5 | 10xK, BSA | 1ul 1ul | 9.0 |
08/29
PCR
Ohara
lane | Primer Foward | Primer Revese | Template |
---|
1 | IK27 | IK28 | Rev, GPDp, GPDt |
2 | IK27 | IK28 | Rev+GPDp, GPDt |
3 | IK27 | IK28 | Rev plasmid |
4 | IK27 | IK28 | Rev plasmid |
5 | IK27 | IK28 | Rev plasmid |
6 | IK27 | IK28 | Rev plasmid |
PCR Polymerase : KAKA Robust
95.0°C : 3min
98.0°C : 20sec
60.0°C : 15sec
72.0°C : 1min30sec
72.0°C : 5min
4.0°C : Infinate
X35 cycle
08/31
Restriction Enzyme digestion
Yoshimoto
lane | Sample DNA name | DNA amount (ul) | RE1 name | RE1 amount(ul) | RE2 name | RE2 amount(ul) | Buffer name | Buffer amount(ul) | Total volume(ul) |
---|
1 | 141 1-2 | 3.5 | EcoR1 | 0.5 | Xba1 | 0.5 | 10xN | 1.0 | 10.0 |
2 | 141 1-2 | 3.5 | EcoR1 | 0.5 | Xba1 | 0.5 | 10xN | 1.0 | 10.0 |
3 | 141 1-3 | 3.5 | EcoR1 | 0.5 | Xba1 | 0.5 | 10xN | 1.0 | 10.0 |
4 | 141 1-3 | 3.5 | EcoR1 | 0.5 | Xba1 | 0.5 | 10xN | 1.0 | 10.0 |
5 | 144-9 | 3.5 | EcoR1 | 0.5 | Not1 | 0.5 | 10xH, BSA | 1ul, 1ul | 10.0 |
6 | 144-9 | 3.5 | EcoR1 | 0.5 | Not1 | 0.5 | 10xH, BSA | 1ul, 1ul | 10.0 |
7 | 144 17 | 3.5 | EcoR1 | 0.5 | Not1 | 0.5 | 10xH, BSA | 1ul, 1ul | 10.0 |
8 | 144 17 | 3.5 | EcoR1 | 0.5 | Not1 | 0.5 | 10xH, BSA | 1ul, 1ul | 10.0 |
9 | 144 3 | 3.5 | EcoR1 | 0.5 | Not1 | 0.5 | 10xH, BSA | 1ul, 1ul | 10.0 |
10 | 144 3 | 3.5 | EcoR1 | 0.5 | Not1 | 0.5 | 10xH, BSA | 1ul, 1ul | 10.0 |
11 | 144 3 | 3.5 | EcoR1 | 0.5 | Not1 | 0.5 | 10xH, BSA | 1ul, 1ul | 10.0 |
12 | 144 1 | 3.5 | EcoR1 | 0.5 | Not1 | 0.5 | 10xH, BSA | 1ul, 1ul | 10.0 |
13 | 144 1 | 3.5 | EcoR1 | 0.5 | Not1 | 0.5 | 10xH, BSA | 1ul, 1ul | 10.0 |
14 | 144 1 | 3.5 | EcoR1 | 0.5 | Not1 | 0.5 | 10xH, BSA | 1ul, 1ul | 10.0 |
15 | 143 21 | 3.5 | Xba1 | 0.5 | Not1 | 0.5 | 10xK, BSA | 1ul, 1ul | 10.0 |
16 | 143 21 | 3.5 | Xba1 | 0.5 | Not1 | 0.5 | 10xK, BSA | 1ul, 1ul | 10.0 |
17 | 143 21 | 3.5 | Xba1 | 0.5 | Not1 | 0.5 | 10xK, BSA | 1ul, 1ul | 10.0 |
18 | 143 15 | 3.5 | Xba1 | 0.5 | Not1 | 0.5 | 10xK, BSA | 1ul, 1ul | 10.0 |
19 | 143 15 | 3.5 | Xba1 | 0.5 | Not1 | 0.5 | 10xK, BSA | 1ul, 1ul | 10.0 |
20 | 143 15 | 3.5 | Xba1 | 0.5 | Not1 | 0.5 | 10xK, BSA | 1ul, 1ul | 10.0 |
21 | 143 15 | 3.5 | Xba1 | 0.5 | Not1 | 0.5 | 10xK, BSA | 1ul, 1ul | 10.0 |
22 | 143 5 | 3.5 | Xba1 | 0.5 | Not1 | 0.5 | 10xK, BSA | 1ul, 1ul | 10.0 |
23 | 143 5 | 3.5 | Xba1 | 0.5 | Not1 | 0.5 | 10xK, BSA | 1ul, 1ul | 10.0 |
24 | 143 5 | 3.5 | Xba1 | 0.5 | Not1 | 0.5 | 10xK, BSA | 1ul, 1ul | 10.0 |
25 | 143 13 | 3.5 | Xba1 | 0.5 | Not1 | 0.5 | 10xK, BSA | 1ul, 1ul | 10.0 |
26 | 145 1-3 | 3.5 | Xba1 | 0.5 | Not1 | 0.5 | 10xK, BSA | 1ul, 1ul | 10.0 |
27 | 145 1-3 | 3.5 | Xba1 | 0.5 | Not1 | 0.5 | 10xK, BSA | 1ul, 1ul | 10.0 |
28 | 145 1-3 | 3.5 | Xba1 | 0.5 | Not1 | 0.5 | 10xK, BSA | 1ul, 1ul | 10.0 |
29 | 142 2-2 | 5.0 | Xba1 | 0.5 | EcoR1 | 0.5 | 10xM | 1.0 | 10.0 |
30 | 142 1-3 | 5.0 | Xba1 | 0.5 | EcoR1 | 0.5 | 10xM | 1.0 | 10.0 |
31 | 142 | 5.0 | Xba1 | 0.5 | EcoR1 | 0.5 | 10xM | 1.0 | 10.0 |
32 | 142 2-2 | 5.0 | Not1 | 0.5 | Xba1 | 0.5 | 10xK BSA | 1ul, 1ul | 10.0 |
33 | 142 1-3 | 5.0 | Not1 | 0.5 | Xba1 | 0.5 | 10xK BSA | 1ul, 1ul | 10.0 |
34 | 142 | 5.0 | Not1 | 0.5 | Xba1 | 0.5 | 10xK BSA | 1ul, 1ul | 10.0 |
09/01
PCR
Yoshimoto
lane | Primer Foward | Primer Revese | Template |
---|
1 | M13F | M13R | P1 |
2 | M13F | M13R | P2 |
3 | M13F | M13R | P3 |
4 | M13F | M13R | P4 |
5 | M13F | M13R | T1 |
6 | M13F | M13R | T2 |
7 | M13F | M13R | T3 |
8 | M13F | M13R | T4 |
PCR Polymerase : KAPA
98.0°C : 3min
98.0°C : 30sec
60.0°C : 20sec
72.0°C : 40sec
72.0°C : 10min
4.0°C : Infinate
X35 cycle
09/02
Restriction Enzyme digestion
Yoshimoto
lane | Sample DNA name | DNA amount (ul) | RE1 name | RE1 amount(ul) | RE2 name | RE2 amount(ul) | Buffer name | Buffer amount(ul) | Total volume(ul) |
---|
1 | 144-3 | 3.5 | Xba1 | 0.5 | EcoR1 | 0.5 | 10xM | 1.0 | 10.0 |
2 | 142 | 3.5 | Xba1 | 0.5 | EcoR1 | 0.5 | 10xM | 1.0 | 10.0 |
3 | 145 1-11 | 3.5 | Xba1 | 0.5 | EcoR1 | 0.5 | 10xM | 1.0 | 10.0 |
09/02
PCR
Yoshimoto
lane | Primer Foward | Primer Revese | Template |
---|
1 | M13F | M13R | 165-1 |
2 | M13F | M13R | 165-2 |
3 | M13F | M13R | 165-3 |
4 | M13F | M13R | 165-4 |
5 | M13F | IK60 | 165-1 |
6 | M13F | IK60 | 165-2 |
7 | M13F | IK60 | 165-3 |
8 | M13F | IK60 | 165-4 |
PCR Polymerase : KAPA
95.0°C : 3min
95.0°C : 30sec
60.0°C : 20sec
72.0°C : 40sec
72.0°C : 5min
4.0°C : Infinate
X35 cycle
09/02
Restriction Enzyme digestion
Yoshimoto
lane | Sample DNA name | DNA amount (ul) | RE1 name | RE1 amount(ul) | RE2 name | RE2 amount(ul) | Buffer name | Buffer amount(ul) | Total volume(ul) |
---|
1 | 142 2-2 | 3.5 | Xba1 | 0.5 | EcoR1 | 0.5 | 10xM | 1.0 | 10.0 |
2 | 1421 | 3.5 | Xba1 | 0.5 | EcoR1 | 0.5 | 10xM | 1.0 | 10.0 |
3 | 142 1-3 | 3.5 | Xba1 | 0.5 | EcoR1 | 0.5 | 10xM | 1.0 | 10.0 |
09/03
Ligation
Yoshimoto
sample | Vector name | Vector amount(ul) | Insert name | Insert amount(ul) |
---|
1 | YEplac195/sma1 | 0.35 | 161 | 6.19 |
2 | YEplac195/sma1 | 0.35 | 162 | 12.2 |
3 | YEplac195/sma1 | 0.35 | 163 | 13.1 |
4 | YEplac195/sma1 | 0.35 | 164 | 10.32 |
5 | YEplac195/sma1 | 0.35 | 166 | 17.25 |
6 | YEplac195/sma1 | 0.35 | 167 | 13.28 |
09/03
PCR
Yoshimoto
lane | Primer Foward | Primer Revese | Template |
---|
1 | M13F | IK55 | GFPgalp-1 |
2 | M13F | IK55 | GFPgalp-2 |
3 | M13F | IK55 | GFPgalp-3 |
4 | M13F | IK55 | GFPgalp-4 |
5 | M13F | IK55 | GFPt-1 |
6 | M13F | IK55 | GFPt-2 |
7 | M13F | IK55 | GFPt-3 |
8 | M13F | IK55 | GFPt-4 |
PCR Polymerase : KAPA
95.0°C : 3min
95.0°C : 30sec
60.0°C : 20sec
72.0°C : 40sec
72.0°C : 5min
4.0°C : Infinate
X35 cycle
09/04
PCR
Ohara
lane | Primer Foward | Primer Revese | Template |
---|
1 | IK21 | IK22 | GPDp |
2 | IK21 | IK22 | GPDp |
3 | IK21 | IK22 | GPDp |
4 | IK21 | IK22 | GPDp |
5 | IK23 | IK28 | GPDter |
6 | IK23 | IK28 | GPDter |
7 | IK23 | IK28 | GPDter |
8 | IK23 | IK28 | GPDter |
9 | IK29 | IK30 | Rev long |
10 | IK29 | IK30 | Rev long |
11 | IK29 | IK30 | Rev long |
12 | IK29 | IK30 | Rev long |
13 | IK21 | IK28 | GPDter(PCRp), Rev(PCRp), GPDp(gel ex) |
14 | IK21 | IK28 | GPDter(PCRp), Rev+GPDp(PCRp) |
PCR Polymerase : KAPA
98.0°C : 3min
98.0°C : 20sec
60.0°C : 30sec
72.0°C : 1min20sec
72.0°C : 5min
4.0°C : Infinate
X35 cycle
PCR pulification: 1~4
PCR pulification: 9~12
Gel extraction: 1~4
Gel extraction: 9~12
09/04
PCR
Yoshimoto
lane | Primer Foward | Primer Revese | Template |
---|
1 | IK60 | AK1F | 142-2-1 |
2 | IK60 | AK1F | 142-2-2 |
3 | IK60 | AK1F | 142-2-3 |
4 | IK60 | AK1F | 142-2-4 |
5 | IK60 | AK1F | 142-2-5 |
6 | IK60 | AK1F | 142-1-1 |
7 | IK60 | AK1F | 142-1-2 |
8 | IK60 | AK1F | 142-1-3 |
9 | IK60 | AK1F | 142-1-4 |
10 | IK60 | AK1F | 142-1-5 |
11 | IK60 | M13F | 144-3 |
12 | IK60 | M13R | 145-1-11 |
13 | IK60 | AK1F | 144-3 |
14 | IK60 | AK1F | 145-1-11 |
PCR Polymerase : KAPA
95.0°C : 3min
95.0°C : 30sec
60.0°C : 20sec
72.0°C : 40sec
72.0°C : 5min
4.0°C : Infinate
X35 cycle
09/04
PCR
Yoshimoto
lane | Primer Foward | Primer Revese | Template |
---|
1 | IK60 | M13R | 142-2-1 |
2 | IK60 | M13R | 142-2-2 |
3 | IK60 | M13R | 142-2-3 |
4 | IK60 | M13R | 142-2-4 |
5 | IK60 | M13R | 142-2-5 |
6 | IK60 | M13R | 142-1-1 |
7 | IK60 | M13R | 142-1-2 |
8 | IK60 | M13R | 142-1-3 |
9 | IK60 | M13R | 142-1-4 |
10 | IK60 | M13R | 142-1-5 |
11 | M13F | M13R | gfpp |
12 | IK55 | M13R | gfpp |
13 | M13F | M13R | gfpt M13 |
14 | IK55 | M13R | gfp 55 |
PCR Polymerase : KAPA
95.0°C : 3min
95.0°C : 20sec
61.0°C : 20sec
72.0°C : 40sec
72.0°C : 5min
4.0°C : Infinate
X35 cycle
09/04
PCR
Ohara
lane | Primer Foward | Primer Revese | Template |
---|
1 | IK27 | IK30 | Rev+p |
2 | IK21 | IK30 | Rev+p |
3 | IK27 | IK22 | GPDp |
4 | IK27 | IK22 | GPDp |
5 | IK27 | IK22 | GPDp |
6 | IK21 | IK22 | GPDp |
7 | IK21 | IK22 | GPDp |
8 | IK21 | IK22 | GPDp |
PCR Polymerase : KAPA
95.0°C : 3min
95.0°C : 15sec
60.0°C : 15sec
72.0°C : 30sec
72.0°C : 2min
4.0°C : Infinate
X35 cycle
09/04
PCR
Fukuda
lane | Primer Foward | Primer Revese | Template |
---|
1 | eef-1g F | eef-1gR | 3 |
2 | eef-1g F | eef-1gR | 4 |
3 | AK1F | AK1R | 5 |
4 | AK1F | AK1R | 6 |
5 | asbF | asbR | 7 |
6 | asbF | asbR | 8 |
PCR Polymerase : KAPA Fast
95.0°C : 3min
95.0°C : 15sec
60.0°C : 15sec
72.0°C : 12sec
72.0°C : 48sec
4.0°C : Infinate
X35 cycle
09/05
PCR
Yoshimoto
lane | Primer Foward | Primer Revese | Template |
---|
1 | M13F | M13R | 161 |
2 | M13F | M13R | 161 |
3 | M13F | M13R | 162 |
4 | M13F | M13R | 163 |
5 | M13F | M13R | 166 |
6 | M13F | M13R | 167 |
7 | M13F | M13R | 168 |
8 | M13F | M13R | 142 |
9 | IK60 | M13R | 161 |
10 | IK60 | M13R | 161 |
11 | M13F | IK60 | 162 |
12 | IK60 | M13R | 163 |
13 | IK60 | M13R | 166 |
14 | M13F | IK60 | 167 |
15 | M13F | IK60 | 168 |
16 | M13F | IK60 | 142 |
PCR Polymerase : KAPA
95.0°C : 3min
95.0°C : 20sec
61.0°C : 20sec
72.0°C : 40sec
72.0°C : 5min
4.0°C : Infinate
X35 cycle
09/05
Restriction Enzyme digestion
Yoshimto
lane | Sample DNA name | DNA amount (ul) | RE1 name | RE1 amount(ul) | RE2 name | RE2 amount(ul) | Buffer name | Buffer amount(ul) | Total volume(ul) |
---|
1 | gfpp | 5.0 | EcoRI | 2.0 | None | None | H | 1.0 | 10.0 |
2 | gfpp | 5.0 | Not1 | 1.0 | HindⅢ | 1.0 | K | 1.0 | 10.0 |
09/05
PCR
Ohara
lane | Primer Foward | Primer Revese | Template |
---|
1 | IK21 | IK28 | Rev, p, t |
2 | IK21 | IK28 | Rev+p, t |
3 | IK27 | IK28 | Rev, p, t |
4 | IK27 | IK28 | Rev+p, t |
5 | IK21 | IK24 | Rev, p, t |
6 | IK21 | IK24 | Rev+p, t |
PCR Polymerase : KAPA
95.0°C : 3min
95.0°C : 15sec
55.0°C : 15sec
72.0°C : 30sec
72.0°C : 2min
4.0°C : Infinate
X35 cycle
09/05
PCR
Ohara
lane | Primer Foward | Primer Revese | Template |
---|
1 | IK21 | IK30 | 9/5 PCR p Rev,pro |
2 | IK21 | IK30 | 9/5 PCR p Rev,pro |
3 | IK21 | IK30 | 9/5 PCR p Rev,pro |
4 | IK21 | IK30 | 9/5 PCR p Rev,pro |
5 | IK29 | IK24 | 9/5 PCR p Rev,ter |
6 | IK29 | IK24 | 9/5 PCR p Rev,ter |
7 | IK29 | IK24 | 9/5 PCR p Rev,ter |
8 | IK29 | IK24 | 9/5 PCR p Rev,ter |
PCR Polymerase : KAPA
95.0°C : 3min
95.0°C : 15sec
55.0°C : 15sec
72.0°C : 30sec
72.0°C : 2min
4.0°C : Infinate
X35 cycle
PCR pulification: 5.0
Gel extraction: 5.0
09/05
PCR
Ohara
lane | Primer Foward | Primer Revese | Template |
---|
1 | IK21 | IK22 | GPD1promoter |
2 | IK21 | IK22 | GPD1promoter |
3 | IK21 | IK22 | GPD1promoter |
4 | IK21 | IK22 | GPD1promoter |
5 | IK21 | IK22 | GPD1promoter |
PCR Polymerase : 2*KAPA Robust
98.0°C : 3min
98.0°C : 20sec
60.0°C : 30sec
72.0°C : 1min
72.0°C : 5min
4.0°C : Infinate
X35 cycle
09/05
PCR
Fukuda
lane | Primer Foward | Primer Revese | Template |
---|
1 | M13F | M13R | 9 |
2 | M13F | M13R | 10 |
3 | 14-3-3 zeta F | 14-3-3 zeta R | 9 |
4 | 14-3-3 zeta F | 14-3-3 zeta R | 10 |
5 | tropomyosin F | tropomyosin R | 11 |
6 | tropomyosin F | tropomyosin R | 12 |
PCR Polymerase : KAPA Fast
95.0°C : 3min
95.0°C : 15sec
60.0°C : 15sec
72.0°C : 12sec
72.0°C : 48sec
4.0°C : Infinate
X35 cycle
09/05
PCR
Fukuda
lane | Primer Foward | Primer Revese | Template |
---|
1 | tropomyosin F | tropomyosin R | 11 |
2 | tropomyosin F | tropomyosin R | 11 |
3 | tropomyosin F | tropomyosin R | 11 |
4 | tropomyosin F | tropomyosin R | 12 |
5 | tropomyosin F | tropomyosin R | 12 |
6 | tropomyosin F | tropomyosin R | 12 |
7 | 14-3-3 pro F | 14-3-3 pro R | 13 |
8 | 14-3-3 pro F | 14-3-3 pro R | 13 |
9 | 14-3-3 pro F | 14-3-3 pro R | 13 |
10 | 14-3-3 pro F | 14-3-3 pro R | 14 |
11 | 14-3-3 pro F | 14-3-3 pro R | 14 |
12 | 14-3-3 pro F | 14-3-3 pro R | 14 |
PCR Polymerase : KAPA Fast
95.0°C : 3min
95.0°C : 15sec
60.0°C : 15sec
72.0°C : 12sec
72.0°C : 48sec
4.0°C : Infinate
X35 cycle
09/05
PCR
Fukuda
lane | Primer Foward | Primer Revese | Template |
---|
1 | tropomyosin F | tropomyosin R | 11 |
2 | tropomyosin F | tropomyosin R | 11 |
3 | tropomyosin F | tropomyosin R | 11 |
4 | tropomyosin F | tropomyosin R | 12 |
5 | tropomyosin F | tropomyosin R | 12 |
6 | tropomyosin F | tropomyosin R | 12 |
7 | eef-1g F | eef-1g R | 4 |
8 | AK1 F | AK1 R | 5 |
9 | M13F | M13R | 9 |
10 | M13F | M13R | 9 |
11 | M13F | M13R | 9 |
12 | M13F | M13R | 9 |
13 | M13F | M13R | 10 |
14 | M13F | M13R | 10 |
15 | M13F | M13R | 10 |
16 | M13F | M13R | 10 |
PCR Polymerase : KAPA Fast
95.0°C : 3min
95.0°C : 15sec
60.0°C : 15sec
72.0°C : 12sec
72.0°C : 48sec
4.0°C : Infinate
X35 cycle
09/06
Restriction Enzyme digestion
Yoshimoto
lane | Sample DNA name | DNA amount (ul) | RE1 name | RE1 amount(ul) | RE2 name | RE2 amount(ul) | Buffer name | Buffer amount(ul) | Total volume(ul) |
---|
1 | gfpp | 5.0 | EcoRI | 2.0 | None | None | H | 1.0 | 10.0 |
2 | gfpp | 5.0 | Not1 | 1.0 | HindⅢ | 1.0 | K | 1.0 | 10.0 |
09/06
PCR
Yoshimoto
lane | Primer Foward | Primer Revese | Template |
---|
1 | M13F | M13R | 163-1 |
2 | M13F | M13R | 163-2 |
3 | M13F | M13R | 164 |
4 | M13F | M13R | 142 |
PCR Polymerase : KAPA
95.0°C : 3min
95.0°C : 20sec
61.0°C : 20sec
72.0°C : 40sec
72.0°C : 5min
4.0°C : Infinate
X35 cycle
09/06
PCR
Ohara
lane | Primer Foward | Primer Revese | Template |
---|
1 | IK29 | IK24 | Rev+t |
2 | IK29 | IK24 | Rev+t |
3 | IK29 | IK24 | Rev+t |
4 | IK29 | IK24 | Rev+t |
5 | IK21 | IK24 | Rev+t, pro PCRp |
6 | IK29 | IK30 | Rev7(plasmid) |
7 | IK29 | IK30 | Rev7(plasmid) |
8 | IK29 | IK30 | Rev7(plasmid) |
PCR Polymerase : KAPA
95.0°C : 3min
95.0°C : 15sec
55.0°C : 15sec
72.0°C : 30sec
72.0°C : 2min
4.0°C : Infinate
X35 cycle
09/06
PCR
Dou
lane | Primer Foward | Primer Revese | Template |
---|
1 | IK21 | IK22 | P1 |
2 | IK21 | IK22 | P1 |
3 | IK21 | IK22 | P1 |
4 | IK21 | IK22 | P1 |
PCR Polymerase : 2X KAPA Robust
98.0°C : 3min
98.0°C : 20sec
60.0°C : 30sec
72.0°C : 1min
72.0°C : 5min
4.0°C : Infinate
X35 cycle
09/07
PCR
Dou
lane | Primer Foward | Primer Revese | Template |
---|
1 | IK21 | IK22 | p+Rev |
2 | IK21 | IK22 | p+Rev |
PCR Polymerase : 2X KAPA Robust
95.0°C : 3min
95.0°C : 15sec
55.0°C : 15sec
72.0°C : 1min
72.0°C : 1min
4.0°C : Infinate
X35 cycle
09/07
PCR
Yoshimoto
lane | Primer Foward | Primer Revese | Template |
---|
1 | IK60 | M13R | 142 PCR |
2 | IK60 | M13R | 142-1 |
3 | IK60 | M13R | 142-2 |
4 | IK60 | M13R | 142-3 |
5 | IK60 | M13R | 142-4 |
6 | IK60 | M13R | 142-5 |
PCR Polymerase : KAPA
95.0°C : 3min
95.0°C : 15sec
60.0°C : 15sec
72.0°C : 40sec
72.0°C : 5min
4.0°C : Infinate
X35 cycle
09/07
PCR
Fukuda
lane | Primer Foward | Primer Revese | Template |
---|
1 | IK61 | IK62 | K733018 |
2 | IK61 | IK62 | K731250 |
PCR Polymerase : KAPA Robust
95.0°C : 3min
95.0°C : 15sec
60.0°C : 15sec
72.0°C : 15sec
72.0°C : 3min
4.0°C : Infinate
X35 cycle
09/07
PCR
Dou
lane | Primer Foward | Primer Revese | Template |
---|
1 | IK21 | IK22 | P1 |
2 | IK21 | IK22 | P1 |
3 | IK21 | IK22 | P1 |
PCR Polymerase : 2X KAPA Robust
98.0°C : 3min
98.0°C : 20sec
60.0°C : 30sec
72.0°C : 1min
72.0°C : 5min
4.0°C : Infinate
X35 cycle
09/08
Restriction Enzyme digestion
Yoshimoto
lane | Sample DNA name | DNA amount (ul) | RE1 name | RE1 amount(ul) | RE2 name | RE2 amount(ul) | Buffer name | Buffer amount(ul) | Total volume(ul) |
---|
1 | 141 | 5.0 | XbaI | 1.0 | NotI | 0.5 | K | 1.0 | 10.0 |
2 | 144 | 5.0 | XbaI | 1.0 | NotI | 0.5 | K | 1.0 | 10.0 |
3 | 143 | 5.0 | XbaI | 1.0 | NotI | 0.5 | K | 1.0 | 10.0 |
4 | 145 | 5.0 | XbaI | 1.0 | NotI | 0.5 | K | 1.0 | 10.0 |
09/08
PCR
Ohara
lane | Primer Foward | Primer Revese | Template |
---|
1 | IK23 | IK28 | Rev3 |
2 | IK23 | IK28 | Rev4 |
3 | IK23 | IK28 | 2(nega) |
4 | IK23 | IK28 | Rev new |
5 | IK29 | IK30 | Rev3 |
6 | IK29 | IK30 | Rev4 |
7 | IK29 | IK30 | 2(nega) |
8 | IK29 | IK30 | Rev new |
PCR Polymerase : KAPA Robust
98.0°C : 3min
98.0°C : 20sec
60.0°C : 30sec
72.0°C : 1min
72.0°C : 5min
4.0°C : Infinate
X35 cycle
09/08
PCR
Dou
lane | Primer Foward | Primer Revese | Template |
---|
1 | IK21 | IK22 | p(blue font) |
2 | IK21 | IK22 | p(blue font) |
3 | IK21 | IK22 | p(blue font) |
4 | IK21 | IK22 | p(blue font) |
PCR Polymerase : 2X KAPA Robust
98.0°C : 3min
98.0°C : 20sec
60.0°C : 30sec
72.0°C : 1min
72.0°C : 5min
4.0°C : Infinate
X35 cycle
09/08
PCR
Ohara
lane | Primer Foward | Primer Revese | Template |
---|
1 | IK23 | IK24 | 99(nega) |
2 | IK23 | IK24 | 101 |
3 | IK23 | IK24 | Rev3 |
4 | IK23 | IK24 | Rev4 |
5 | IK23 | IK24 | Rev5 |
6 | IK23 | IK24 | Rev6 |
7 | IK23 | IK24 | T(posi) |
8 | IK29 | IK30 | 99 |
9 | IK29 | IK30 | Rev5 |
10 | IK29 | IK30 | Rev6 |
PCR Polymerase : KAPA
95.0°C : 3min
95.0°C : 15sec
55.0°C : 15sec
72.0°C : 30sec
72.0°C : 2min
4.0°C : Infinate
X35 cycle
09/08
PCR
Yoshimoto
lane | Primer Foward | Primer Revese | Template |
---|
1 | IK21 | IK24 | all |
2 | IK21 | IK24 | all |
3 | IK21 | IK22 | promoter |
4 | IK21 | IK22 | promoter |
5 | IK21 | IK22 | promoter |
6 | IK21 | IK22 | promoter |
7 | IK29 | IK30 | Rev |
8 | IK29 | IK30 | Rev |
9 | IK29 | IK30 | nega 99 |
10 | IK29 | IK30 | water |
11 | IK29 | IK24 | Rev+t |
12 | IK29 | IK24 | Rev+t |
13 | IK23 | IK24 | plasmid |
14 | IK23 | IK24 | nega 99 |
15 | IK23 | IK24 | posi(T PCRp) |
16 | IK23 | IK24 | posi(T PCRp) |
17 | IK23 | IK24 | posi(T PCRp) |
18 | IK23 | IK24 | posi(T PCRp) |
19 | IK23 | IK24 | water |
PCR Polymerase : go taq
95.0°C : 2min
95.0°C : 1min
55.0°C : 1min
72.0°C : 1min
72.0°C : 5min
10.0°C : Infinate
X35 cycle
09/08
PCR
Yoshimoto
lane | Primer Foward | Primer Revese | Template |
---|
1 | IK21 | IK22 | PCRp X10 |
2 | IK21 | IK22 | PCRp X10 |
3 | IK21 | IK22 | genome X10 |
4 | IK21 | IK22 | genome |
5 | IK21 | IK22 | water |
6 | IK21 | IK24 | all |
7 | IK21 | IK24 | p(PCRp),Rev+t gel ex |
8 | IK21 | IK24 | 8/20 PCRp,Rev+t gel ex |
9 | IK21 | IK24 | RA IK21X10 |
10 | IK21 | IK24 | RA |
11 | IK21 | IK24 | water |
PCR Polymerase : go taq
95.0°C : 2min
95.0°C : 1min
55.0°C : 1min
72.0°C : 2min
72.0°C : 5min
10.0°C : Infinate
X35 cycle
09/09
Ligation
Yoshimoto
sample | Vector name | Vector amount(ul) | Insert name | Insert amount(ul) |
---|
1 | 143 | 2 | 141.0 | 2.0 |
2 | 145 | 2 | 144.0 | 2.0 |
3 | 143 | 2 | 144.0 | 2.0 |
09/09
Restriction Enzyme digestion
Yoshimoto
lane | Sample DNA name | DNA amount (ul) | RE1 name | RE1 amount(ul) | RE2 name | RE2 amount(ul) | Buffer name | Buffer amount(ul) | Total volume(ul) |
---|
1 | 142 | 20.0 | XbaI | 4.0 | NotI | 2.0 | K | 4.0 | 40.0 |
09/09
Ligation
Yoshimoto
sample | Vector name | Vector amount(ul) | Insert name | Insert amount(ul) |
---|
1 | 323 | 0.34 | GFPp(gal1) | 12.5 |
09/10
Restriction Enzyme digestion
Yoshimoto
lane | Sample DNA name | DNA amount (ul) | RE1 name | RE1 amount(ul) | RE2 name | RE2 amount(ul) | Buffer name | Buffer amount(ul) | Total volume(ul) |
---|
1 | 142-3 | 5.0 | XbaI | 0.5 | NotI | 0.5 | K | 1.0 | 10.0 |
2 | 142-5 | 5.0 | XbaI | 0.5 | NotI | 0.5 | K | 1.0 | 10.0 |
3 | 161-1 | 5.0 | XbaI | 0.5 | NotI | 0.5 | K | 1.0 | 10.0 |
4 | 163-1 | 5.0 | XbaI | 0.5 | NotI | 0.5 | K | 1.0 | 10.0 |
5 | 164 | 5.0 | XbaI | 0.5 | NotI | 0.5 | K | 1.0 | 10.0 |
6 | 166 | 5.0 | XbaI | 0.5 | NotI | 0.5 | K | 1.0 | 10.0 |
09/14
PCR
Yoshimoto
lane | Primer Foward | Primer Revese | Template |
---|
1 | M13F | IK60 | 162 |
2 | IK29 | IK24 | Rev |
3 | IK40 | IK55 | GFP |
PCR Polymerase : go taq
95.0°C : 2min
95.0°C : 1min
58.0°C : 1min
72.0°C : 1min
72.0°C : 2min
10.0°C : Infinate
X35 cycle
09/14
Ligation
Yoshimoto
sample | Vector name | Vector amount(ul) | Insert name | Insert amount(ul) |
---|
1 | YEplac195/sma1 | 0.34 | 163.0 | 13.1 |
2 | YEplac195/sma1 | 0.34 | 166.0 | 17.25 |
3 | 143 | 2 | 141.0 | 2.0 |
4 | 145 | 2 | 144.0 | 2.0 |
5 | 143 | 2 | 144.0 | 2.0 |
09/14
PCR
Ohara
lane | Primer Foward | Primer Revese | Template |
---|
1 | IK21 | IK22 | genome |
2 | IK21 | IK22 | genome |
3 | IK121 | IK122 | genome |
4 | IK121 | IK122 | genome |
5 | IK129 | IK130 | Rev |
6 | IK129 | IK130 | Rev |
7 | IK129 | IK130 | nega 99 |
8 | IK129 | IK130 | water |
9 | IK21 | IK22 | water |
10 | IK121 | IK122 | water |
PCR Polymerase : 2X primestar
98.0°C : 3min
98.0°C : 10sec
55.0°C : 15sec
72.0°C : 5sec
72.0°C : 3min
4.0°C : Infinate
X35 cycle
PCR pulification: 1,2
PCR pulification: 4,5
Gel extraction: 1,2
Gel extraction: 4,5
09/14
PCR
Ohara
lane | Primer Foward | Primer Revese | Template |
---|
1 | IK121 | IK122 | p |
2 | IK121 | IK122 | p |
3 | IK21 | IK22 | p |
4 | IK21 | IK22 | p |
5 | IK121 | IK122 | genome |
6 | IK121 | IK122 | genome |
7 | IK21 | IK24 | p1, Rev+t |
8 | IK21 | IK24 | p1, Rev+t |
9 | IK129 | IK24 | Rev new, t |
10 | IK21 | IK130 | p, Rev new |
11 | IK129 | IK24 | water |
12 | IK21 | IK130 | water |
PCR Polymerase : prime star max
98.0°C : 3min
98.0°C : 10sec
55.0°C : 15sec
72.0°C : 5sec
72.0°C : 3min
4.0°C : Infinate
X35 cycle
09/15
PCR
Ohara
lane | Primer Foward | Primer Revese | Template |
---|
1 | IK129 | IK130 | Rev |
2 | IK129 | IK130 | Rev |
3 | IK129 | IK130 | Rev |
4 | IK21 | IK24 | OP2+(Rev+t) |
5 | IK129 | IK24 | NR1+t |
6 | IK21 | IK130 | NR1+OP2 |
PCR Polymerase : primestar
98.0°C : 3min
98.0°C : 10sec
55.0°C : 5sec
72.0°C : 1min30sec
72.0°C : 3min
4.0°C : Infinate
X35 cycle
09/15
PCR
Ohara
lane | Primer Foward | Primer Revese | Template |
---|
1 | IK129 | IK24 | NR1+t |
2 | IK129 | IK24 | NR1+t |
3 | IK129 | IK24 | NR1+t |
4 | IK129 | IK24 | NR1+t |
5 | IK129 | IK24 | water |
6 | K129 | IK130 | rev |
7 | K129 | IK130 | rev |
PCR Polymerase : primestar
98.0°C : 3min
98.0°C : 10sec
55.0°C : 5sec
72.0°C : 1min30sec
72.0°C : 3min
4.0°C : Infinate
X35 cycle
PCR pulification: 1~5
PCR pulification: 6,7
Gel extraction: 1~5
Gel extraction: 6,7
09/15
PCR
Ohara
lane | Primer Foward | Primer Revese | Template |
---|
1 | IK129 | IK24 | RT1 |
2 | IK121 | IK122 | genome |
3 | IK121 | IK122 | genome |
4 | IK121 | IK122 | genome |
5 | IK21 | IK24 | RT1+OP1 |
6 | IK21 | IK24 | RT1+OP2 |
7 | IK121 | IK24 | RT1+NP1 |
PCR Polymerase : primestar
98.0°C : 3min
98.0°C : 10sec
55.0°C : 5sec
72.0°C : 1min30sec
72.0°C : 3min
4.0°C : Infinate
X35 cycle
09/15
PCR
Ohara
lane | Primer Foward | Primer Revese | Template |
---|
1 | IK121 | IK24 | RT1+NP1 |
2 | IK121 | IK24 | RT1+NP1 |
3 | IK121 | IK24 | RT1+NP1 |
4 | IK121 | IK24 | RT1+NP1 |
5 | IK129 | IK24 | RT1 |
6 | IK121 | IK24 | water |
PCR Polymerase : primestar
98.0°C : 3min
98.0°C : 10sec
55.0°C : 5sec
72.0°C : 1min30sec
72.0°C : 3min
4.0°C : Infinate
X35 cycle
PCR pulification: 1~4
Gel extraction: 1~4
09/16
PCR
Yoshimoto
lane | Primer Foward | Primer Revese | Template |
---|
1 | IK60 | M13R | 161-1 |
2 | IK60 | M13R | 161-2 |
3 | IK60 | M13R | 163-1 |
4 | IK60 | M13R | 163-2 |
5 | IK60 | M13R | 163-3 |
6 | IK60 | M13R | 163-4 |
7 | IK60 | M13R | 166-1 |
8 | IK60 | M13R | 166-2 |
9 | IK60 | M13R | 166-3 |
10 | IK60 | M13R | 166-4 |
11 | IK60 | M13R | 166-5 |
12 | IK60 | M13R | 166-6 |
13 | IK60 | M13R | 162-1 |
14 | IK60 | M13R | 162-3 |
15 | IK60 | M13R | 161-1 |
16 | IK60 | M13R | 165-2 |
17 | IK60 | M13R | 165-3 |
18 | IK60 | M13R | 165-4 |
19 | IK60 | M13R | 165-5 |
20 | IK60 | M13R | 167-1 |
21 | IK60 | M13R | 167-2 |
22 | IK60 | M13R | 167-3 |
23 | M13R | IK40 | Gal1GFP 1 |
24 | M13R | IK40 | Gal1GFP 2 |
25 | M13R | IK40 | Gal1GFP 3 |
26 | M13R | IK40 | Gal1GFP 4 |
PCR Polymerase : go taq
95.0°C : 2min
95.0°C : 1min
60.0°C : 1min
72.0°C : 2min
72.0°C : 5min
4.0°C : Infinate
X35 cycle
09/16
Ligation
Ohara
sample | Vector name | Vector amount(ul) | Insert name | Insert amount(ul) |
---|
1 | 325 | 0.528 | prt2 | 0.26 |
2 | 325 | 3.65 | prt2 | 7.33 |
09/16
PCR
Ohara
lane | Primer Foward | Primer Revese | Template |
---|
1 | IK121 | IK24 | prt2 |
2 | IK121 | IK24 | prt3 |
3 | IK121 | IK24 | prt3 |
4 | IK121 | IK24 | prt3 |
5 | IK121 | IK24 | prt3 |
6 | IK121 | IK122 | genome |
PCR Polymerase : primestar
98.0°C : 3min
98.0°C : 10sec
55.0°C : 5sec
72.0°C : 10sec
72.0°C : 2min
4.0°C : Infinate
X35 cycle
PCR pulification: 2~5
Gel extraction: 2~5
09/16
PCR
Yoshimoto
lane | Primer Foward | Primer Revese | Template |
---|
1 | IK60 | M13R | 142-5 |
2 | IK60 | M13R | 142-5 |
3 | M13F | IK60 | 143-1 |
4 | M13F | IK60 | 143-2 |
5 | M13F | IK60 | 143-3 |
6 | M13F | IK60 | 145-1 |
7 | M13F | IK60 | 145-2 |
8 | M13F | IK60 | 145-3 |
PCR Polymerase : go taq
95.0°C : 2min
95.0°C : 1min
60.0°C : 1min
72.0°C : 2min
72.0°C : 5min
4.0°C : Infinate
X35 cycle
09/16
PCR
Ohara
lane | Primer Foward | Primer Revese | Template |
---|
1 | M13F | M13R | genome |
PCR Polymerase : go taq
95.0°C : 2min
95.0°C : 1min
60.0°C : 1min
72.0°C : 2min
72.0°C : 5min
4.0°C : Infinate
X35 cycle
09/16
PCR
Yoshimoto
lane | Primer Foward | Primer Revese | Template |
---|
1 | IK60 | M13F | 164-1 |
2 | IK60 | M13F | 164-2 |
3 | M13F | IK40 | Gal1 GFP 1 |
4 | M13F | IK40 | Gal1 GFP 2 |
5 | M13F | IK40 | Gal1 GFP 3 |
6 | M13F | IK40 | Gal1 GFP 4 |
PCR Polymerase : go taq
95.0°C : 2min
95.0°C : 1min
60.0°C : 1min
72.0°C : 1min
72.0°C : 5min
4.0°C : Infinate
X35 cycle
09/17
PCR
Ohara
lane | Primer Foward | Primer Revese | Template |
---|
1 | IK121' | IK24 | N(blue colony) |
2 | IK121' | IK24 | 1(Rev white) |
3 | IK121' | IK24 | 2 |
4 | IK121' | IK24 | 3 |
5 | IK121' | IK24 | prt3 |
6 | IK121' | IK122' | genome |
PCR Polymerase : prime star
98.0°C : 3min
98.0°C : 10sec
55.0°C : 5sec
72.0°C : 10sec
72.0°C : 2min
4.0°C : Infinate
X35 cycle
09/17
PCR
Ohara
lane | Primer Foward | Primer Revese | Template |
---|
1 | M13F | M13R | 172-0 |
2 | M13F | M13R | 172-1 |
3 | M13F | M13R | 172-2 |
4 | M13F | M13R | 172-3 |
5 | M13F | M13R | 172-4 |
6 | M13F | M13R | 172-5 |
7 | M13F | M13R | 172-6 |
8 | M13F | M13R | 172-8 |
9 | M13F | M13R | 172-9 |
10 | M13F | M13R | 172-10 |
11 | M13F | M13R | 172-11 |
12 | M13F | M13R | 172-12 |
13 | M13F | M13R | 172-13 |
14 | M13F | M13R | 114-11 |
15 | M13F | M13R | 114-13 |
16 | M13F | M13R | 114-0 |
17 | M13F | M13R | 171-1-1 |
18 | M13F | M13R | 171-2 |
19 | M13F | M13R | 174-1 |
20 | M13F | M13R | 175-1 |
21 | M13F | M13R | 175-2 |
22 | M13F | M13R | 175-3 |
23 | M13F | M13R | 175-4 |
24 | M13F | M13R | 176-0 |
25 | M13F | M13R | 176-1 |
26 | M13F | M13R | 176-2 |
27 | M13F | M13R | 177-1 |
28 | M13F | M13R | 177-2 |
29 | M13F | M13R | 177-3 |
30 | M13F | M13R | 177-4 |
31 | M13F | M13R | 177-5 |
32 | M13F | M13R | water |
33 | M13F | M13R | 26(GPD AK1) |
PCR Polymerase : go taq
95.0°C : 2min15sec
95.0°C : 1min
55.0°C : 1min
72.0°C : 2min
72.0°C : 5min
4.0°C : Infinate
X35 cycle
09/17
Ligation
Ohara
sample | Vector name | Vector amount(ul) | Insert name | Insert amount(ul) |
---|
1 | 325 | 0.528 | prt4 | 11.7 |
09/18
PCR
Yoshimoto
lane | Primer Foward | Primer Revese | Template |
---|
1 | IK40 | IK47 | 114-0 |
2 | IK55 | IK40 | 171-1 |
3 | IK40 | IK41 | 172-1 |
4 | IK60 | AK1F | 176-0 |
5 | IK60 | AK1F | 177-1 |
6 | IK60 | AK1F | 177-2 |
PCR Polymerase : go taq
95.0°C : 2min15sec
95.0°C : 1min
58.0°C : 1min
72.0°C : 50sec
72.0°C : 5min
4.0°C : Infinate
X35 cycle
09/18
PCR
Yoshimoto
lane | Primer Foward | Primer Revese | Template |
---|
1 | IK60 | M13R | 104-1 |
2 | IK60 | M13R | 104-2 |
PCR Polymerase : go taq
95.0°C : 2min15sec
95.0°C : 1min
60.0°C : 1min
72.0°C : 5min
72.0°C : 5min
4.0°C : Infinate
X35 cycle
09/19
PCR
Yoshimoto
lane | Primer Foward | Primer Revese | Template |
---|
1 | PrimerF | PrimerR | 162-1 |
2 | PrimerF | PrimerR | 162-1 |
3 | PrimerF | PrimerR | 162-1 |
4 | PrimerF | PrimerR | 162-1 |
5 | PrimerF | PrimerR | Rev |
6 | PrimerF | PrimerR | Rev |
7 | PrimerF | PrimerR | Rev |
8 | PrimerF | PrimerR | Rev |
PCR Polymerase : go taq
95.0°C : 2min15sec
95.0°C : 1min
55.0°C : 1min
72.0°C : 2min15sec
72.0°C : 5min
4.0°C : Infinate
X35 cycle
09/19
PCR
Yoshimoto
lane | Primer Foward | Primer Revese | Template |
---|
1 | Ik29 | IK30 | OR1-1 |
2 | Ik29 | IK30 | OR1-2 |
3 | Ik29 | IK30 | OR1-3 |
4 | Ik29 | IK30 | OR1-4 |
5 | Ik29 | IK30 | OR1 nega |
6 | Ik29 | IK30 | R2 |
PCR Polymerase : go taq
95.0°C : 2min15sec
95.0°C : 1min
55.0°C : 1min
72.0°C : 2min15sec
72.0°C : 5min
4.0°C : Infinate
X35 cycle
09/19
PCR
Yoshimoto
lane | Primer Foward | Primer Revese | Template |
---|
1 | IK60 | M13R | 162-1 |
2 | IK60 | M13R | 162-2 |
3 | IK60 | M13R | 162-3 |
PCR Polymerase : go taq
95.0°C : 2min15sec
95.0°C : 1min
55.0°C : 1min
72.0°C : 2min15sec
72.0°C : 5min
4.0°C : Infinate
X35 cycle
09/19
PCR
Yoshimoto
lane | Primer Foward | Primer Revese | Template |
---|
1 | IK89 | IK90 | EGFP |
2 | IK89 | IK90 | EGFP |
3 | IK84 | IK85 | REV |
4 | IK86 | IK87 | REV |
PCR Polymerase : go taq
95.0°C : 2min15sec
95.0°C : 1min
55.0°C : 1min
72.0°C : 2min15sec
72.0°C : 5min
4.0°C : Infinate
X35 cycle
PCR pulification: lane1,2
PCR pulification: lane3
Gel extraction: lane1,2
Gel extraction: lane3
09/20
PCR
Yoshimoto
lane | Primer Foward | Primer Revese | Template |
---|
1 | IK40 | IK55 | 171 |
2 | IK40 | IK55 | 171 |
3 | IK40 | IK55 | 171 |
4 | IK40 | IK55 | 171 |
5 | IK40 | IK55 | 171 |
6 | IK40 | IK55 | 171 |
7 | IK40 | IK60 | 177 |
8 | IK40 | IK60 | 177 |
9 | IK40 | IK41 | 172 |
10 | IK40 | IK41 | 172 |
11 | IK40 | IK41 | 172 |
12 | IK40 | IK41 | 172 |
13 | IK40 | IK41 | 172 |
14 | IK40 | IK41 | 172 |
15 | IK40 | IK60 | 176 |
16 | IK40 | IK60 | 176 |
17 | IK46 | M13F | 148 |
18 | IK46 | M13F | 148 |
19 | IK46 | M13F | 148 |
20 | IK46 | M13F | 148 |
21 | IK46 | M13F | 148 |
PCR Polymerase : go taq
95.0°C : 2min15sec
95.0°C : 1min
55.0°C : 1min
72.0°C : 2min15sec
72.0°C : 5min
4.0°C : Infinate
X35 cycle
09/21
PCR
Yoshimoto
lane | Primer Foward | Primer Revese | Template |
---|
1 | M13F | M13R | OR-1(REV plasmid) |
2 | M13F | M13R | OR-2 |
3 | M13F | M13R | OR-3 |
4 | M13F | M13R | OR-4 |
5 | M13F | M13R | OR-5 |
6 | IK23 | Ik24 | OR-1 |
7 | IK23 | Ik24 | OR-2 |
8 | IK23 | Ik24 | OR-3 |
9 | IK23 | Ik24 | OR-4 |
10 | IK23 | Ik24 | OR-5 |
PCR Polymerase : go taq
95.0°C : 2min15sec
95.0°C : 1min
55.0°C : 1min
72.0°C : 2min15sec
72.0°C : 5min
4.0°C : Infinate
X35 cycle
09/22
PCR
Ohara
lane | Primer Foward | Primer Revese | Template |
---|
1 | IK121 | IK24 | prt3 |
2 | IK121 | IK24 | num99 |
3 | IK121 | IK24 | OR1-1 |
4 | IK121 | IK24 | OR1-2 |
5 | IK121 | IK24 | OR1-3 |
6 | IK121 | IK24 | OR1-5 |
7 | IK121 | IK24 | OR1-4 |
PCR Polymerase : prime star max
98.0°C : 3min
98.0°C : 10sec
55.0°C : 5sec
72.0°C : 10sec
72.0°C : 2min
4.0°C : Infinate
X35 cycle
09/22
PCR
Dou
lane | Primer Foward | Primer Revese | Template |
---|
1 | IK89 | IK90 | EGFP2-1 |
2 | IK89 | IK90 | EGFP2-2 |
PCR Polymerase : go taq
95.0°C : 2min15sec
95.0°C : 1min
55.0°C : 1min
72.0°C : 2min15sec
72.0°C : 5min
4.0°C : Infinate
X35 cycle
10/04
Restriction Enzyme digestion
yoshimoto
lane | Sample DNA name | DNA amount (ul) | RE1 name | RE1 amount(ul) | RE2 name | RE2 amount(ul) | Buffer name | Buffer amount(ul) | Total volume(ul) |
---|
1 | Template 5 | 5.0 | Xba | 0.025 | EcoR1 | 0.025 | M | 1.0 | 10.0 |
2 | Template 5 | 5.0 | Not1 | 0.025 | EcoR1 | 0.025 | H | 1.0 | 10.0 |
3 | Template 5 | 5.0 | Not1 | 0.025 | Xba1 | 0.025 | H | 1.0 | 10.0 |
10/04
PCR
ohara
lane | Primer Foward | Primer Revese | Template |
---|
1 | OD21 | OD23 | eGFP plasmid |
2 | OD21 | OD23 | eGFP plasmid |
3 | OD22 | OD23 | eGFP plasmid |
4 | OD22 | OD23 | eGFP plasmid |
5 | OD31 | OD32 | T7-RRE c |
6 | OD31 | OD32 | T7-RRE c |
PCR Polymerase : prime star max
98.0°C : 10sec
98.0°C : 10sec
55.0°C : 5sec
72.0°C : 10sec
72.0°C : 10sec
15.0°C : Infinate
X35 cycle
10/04
Restriction Enzyme digestion
Yoshimoto
lane | Sample DNA name | DNA amount (ul) | RE1 name | RE1 amount(ul) | RE2 name | RE2 amount(ul) | Buffer name | Buffer amount(ul) | Total volume(ul) |
---|
1 | pSB1C3 | 15.0 | Xba1 | 5.0 | Spe1 | 5.0 | M | 12.0 | 100.0 |
2 | pSB1C3 | 15.0 | Xba1 | 5.0 | Pst1 | 5.0 | M | 12.0 | 100.0 |
10/05
PCR
yoshimoto
lane | Primer Foward | Primer Revese | Template |
---|
1 | vf | vr | Colony1~20(191) |
2 | vf | vr | Colony21~40(192) |
3 | vf | vr | Colony41~60(193) |
4 | vf | vr | Colony61~80(194) |
PCR Polymerase : prime star max
98.0°C : 10sec
98.0°C : 10sec
55.0°C : 5sec
72.0°C : 10sec
72.0°C : 10sec
15.0°C : Infinate
X35 cycle
10/05
Restriction Enzyme digestion
yoshimoto
lane | Sample DNA name | DNA amount (ul) | RE1 name | RE1 amount(ul) | Buffer name | Buffer amount(ul) | Total volume(ul) |
---|
1 | template | 5.0 | Pst1 | 0.05 | 10xH | 1.0 | 10.0 |
10/06
PCR
yoshimoto
lane | Primer Foward | Primer Revese | Template |
---|
1 | OD41 | OD42 | Gal-GFP |
2 | OD41 | OD43 | Gal-GFP |
3 | OD41 | OD44 | Gal-GFP |
4 | IK61 | OD52 | Gal-GFP |
5 | OD53 | OD54 | Gal-GFP |
PCR Polymerase : prime star max
98.0°C : 10sec
98.0°C : 10sec
55.0°C : 5sec
72.0°C : 10sec
72.0°C : 10sec
15.0°C : Infinate
X35 cycle
10/07
PCR
yoshimoto
lane | Primer Foward | Primer Revese | Template |
---|
1 | IK61 | OD54 | T7-RRE |
2 | IK61 | OD54 | fukuda GFP |
PCR Polymerase : pfu
95.0°C : 2min
95.0°C : 1min
55.0°C : 30sec
72.0°C : 4min
72.0°C : 4min
15.0°C : Infinate
X40 cycle
10/07
PCR
yoshimoto
lane | Primer Foward | Primer Revese | Template |
---|
1 | OD11 | OD12 | GPD-gfp |
2 | OD11 | OD13 | GPD-gfp |
3 | OD41 | OD42 | Gal-GFP |
4 | OD41 | OD43 | Gal-GFP |
5 | OD41 | OD44 | Gal-GFP |
6 | OD31 | OD32 | T7-RRE |
7 | IK61 | IK54 | fukuda GFP + T7RRE |
8 | IK61 | IK54 | fukuda GFP + T7RRE |
PCR Polymerase : pfu
95.0°C : 2min
95.0°C : 1min
55.0°C : 30sec
72.0°C : 4min
72.0°C : 4min
15.0°C : Infinate
X40 cycle
10/09
Restriction Enzyme digestion
yoshimoto
lane | Sample DNA name | DNA amount (ul) | RE1 name | RE1 amount(ul) | RE2 name | RE2 amount(ul) | Buffer name | Buffer amount(ul) | Total volume(ul) |
---|
1 | T7 RRE | 15.0 | Xba1 | 3.0 | Pst1 | 3.0 | M | 3.0 | 30.0 |
10/09
PCR
yoshimoto
lane | Primer Foward | Primer Revese | Template |
---|
1 | IK61 | IK54 | GFP+RRE |
PCR Polymerase : pfu
95.0°C : 2min
95.0°C : 1min
55.0°C : 30sec
72.0°C : 4min
72.0°C : 4min
15.0°C : Infinate
X40 cycle
10/09
PCR
yoshimoto
lane | Primer Foward | Primer Revese | Template |
---|
1 | OD41 | OD42 | Gal-gfp |
2 | OD41 | OD44 | Gal-gfp |
PCR Polymerase : pfu
95.0°C : 2min
95.0°C : 1min
55.0°C : 30sec
72.0°C : 4min
72.0°C : 4min
15.0°C : Infinate
X35 cycle
10/12
PCR
yoshimoto
lane | Primer Foward | Primer Revese | Template |
---|
1 | OD61 | OD12 | TDH3-eGFP |
2 | OD62 | OD12 | TDH3-eGFP |
3 | OD63 | OD12 | TDH3-eGFP |
4 | OD61 | OD13 | TDH3-eGFP |
5 | OD62 | OD13 | TDH3-eGFP |
6 | OD63 | OD13 | TDH3-eGFP |
7 | OD73 | IK62 | GFP |
8 | OD45 | OD42 | GFP |
9 | OD45 | OD43 | GFP |
10 | OD45 | OD44 | GFP |
11 | IK61 | OD74 | GFP |
PCR Polymerase : KAPA HiFi fast
95.0°C : 5min
98.0°C : 20sec
58.0°C : 15sec
72.0°C : 40sec
72.0°C : 5min
4.0°C : Infinate
X35 cycle
10/12
Restriction Enzyme digestion
Yoshimoto
lane | Sample DNA name | DNA amount (ul) | RE1 name | RE1 amount(ul) | Buffer name | Buffer amount(ul) | Total volume(ul) |
---|
1 | Template14 | 5.0 | Xba1 | 1.0 | M | 1.0 | 10.0 |
2 | Template100 | 5.0 | Xba1 | 1.0 | M | 1.0 | 10.0 |
10/12
PCR
yoshimoto
lane | Primer Foward | Primer Revese | Template |
---|
1 | OD62 | OD12 | yeast genome |
2 | OD62 | OD13 | yeast genome |
3 | OD73 | IK62 | fukuda gfp |
4 | OD45 | OD42 | gal gfp |
5 | IK61 | OD74 | fukuda gfp |
PCR Polymerase : KAPA hifi
95.0°C : 5min
98.0°C : 20sec
58.0°C : 15sec
72.0°C : 40sec
72.0°C : 5min
4.0°C : Infinate
X35 cycle
10/13
PCR
yoshimoto
lane | Primer Foward | Primer Revese | Template |
---|
1 | OD62 | OD12 | yeast genome |
2 | OD62 | OD12 | yeast genome |
3 | OD62 | OD13 | TDH3-eGFP |
4 | OD62 | OD13 | TDH3-eGFP |
5 | OD73 | IK62 | fukuda GFP |
6 | IK61 | OD71 | fukuda GFP |
7 | OD72 | OD54 | T7-RRE |
PCR Polymerase : KAPA hifi
95.0°C : 5min
98.0°C : 20sec
58.0°C : 15sec
72.0°C : 40sec
72.0°C : 5min
4.0°C : Infinate
X35 cycle
10/14
PCR
yoshimoto
lane | Primer Foward | Primer Revese | Template |
---|
1 | OD62 | OD12 | yeast genome |
2 | OD62 | OD13 | yeast genome |
3 | OD45 | OD42 | gal1 GFP |
4 | IK61 | OD74 | fukuda GFP |
5 | OD73 | IK62 | fukuda GFP |
PCR Polymerase : KAPA
95.0°C : 5min
98.0°C : 20sec
58.0°C : 15sec
72.0°C : 40sec
72.0°C : 5min
4.0°C : Infinate
X35 cycle
10/14
PCR
ohara
lane | Primer Foward | Primer Revese | Template |
---|
1 | IK103 | IK104 | 165 gBlock |
2 | IK107 | IK108 | 161 gBlock |
3 | IK101 | IK102 | 161 gBlock |
4 | IK105 | IK106 | 163 gBlock |
PCR Polymerase : yoshimoto
95.0°C : 5min
98.0°C : 20sec
58.0°C : 15sec
72.0°C : 40sec
72.0°C : 5min
4.0°C : Infinate
X35 cycle
10/14
PCR
yoshimoto
lane | Primer Foward | Primer Revese | Template |
---|
1 | OD45 | OD42 | gal1 GFP |
2 | IK61 | OD74 | fukuda GFP |
3 | OD73 | IK62 | fukuda GFP |
4 | IK61 | OD54 | fukuda GFP |
PCR Polymerase : KAPA hifi
95.0°C : 5min
98.0°C : 20sec
58.0°C : 15sec
72.0°C : 40sec
72.0°C : 5min
4.0°C : Infinate
X35 cycle
10/14
Restriction Enzyme digestion
yoshimoto
lane | Sample DNA name | DNA amount (ul) | RE1 name | RE1 amount(ul) | Buffer name | Buffer amount(ul) | Total volume(ul) |
---|
1 | template | 15.0 | Xba1 | 3.0 | M | 3.0 | 100.0 |
2 | template | 15.0 | Xba1 | 5.0 | M | 3.0 | 100.0 |
10/14
PCR
yoshimoto
lane | Primer Foward | Primer Revese | Template |
---|
1 | OD31 | OD32 | T7+RRE |
2 | OD22 | OD23 | eGFP |
PCR Polymerase : KAPA hifi
95.0°C : 5min
98.0°C : 20sec
58.0°C : 15sec
72.0°C : 40sec
72.0°C : 5min
4.0°C : Infinate
X35 cycle
10/15
PCR
yoshimoto
lane | Primer Foward | Primer Revese | Template |
---|
1 | IK61 | OD54 | 6+7 |
PCR Polymerase : KAPA hifi
95.0°C : 5min
98.0°C : 20sec
58.0°C : 15sec
72.0°C : 40sec
72.0°C : 5min
4.0°C : Infinate
X35 cycle
10/15
PCR
yoshimoto
lane | Primer Foward | Primer Revese | Template |
---|
1 | IK85 | IK86 | rev |
2 | IK87 | Ik88 | rev |
PCR Polymerase : KAPA hifi
95.0°C : 5min
98.0°C : 20sec
58.0°C : 15sec
72.0°C : 40sec
72.0°C : 5min
4.0°C : Infinate
X35 cycle
PCR pulification: 1~2
Gel extraction: 1~2
10/15
PCR
yoshimoto
lane | Primer Foward | Primer Revese | Template |
---|
1 | OD31 | OD32 | T7+RRE |
2 | OD22 | OD23 | TDH3promoter |
PCR Polymerase : KAPA hifi
95.0°C : 5min
98.0°C : 20sec
58.0°C : 15sec
72.0°C : 40sec
72.0°C : 5min
4.0°C : Infinate
X35 cycle
10/15
PCR
yoshimoto
lane | Primer Foward | Primer Revese | Template |
---|
1 | OD31 | OD32 | T7+RRE |
2 | OD22 | OD23 | TDH3promoter |
3 | OD45 | OD42 | Gal eGFP |
PCR Polymerase : KAPA hifi
95.0°C : 5min
98.0°C : 20sec
58.0°C : 15sec
72.0°C : 40sec
72.0°C : 5min
4.0°C : Infinate
X30 cycle
10/15
PCR
yoshimoto
lane | Primer Foward | Primer Revese | Template |
---|
1 | IK105 | Ik106 | Gblock 163 |
PCR Polymerase : KAPA hifi
95.0°C : 5min
98.0°C : 20sec
58.0°C : 15sec
72.0°C : 40sec
72.0°C : 5min
4.0°C : Infinate
X35 cycle
10/15
PCR
yoshimoto
lane | Primer Foward | Primer Revese | Template |
---|
1 | IK105 | IK108 | Gal1 loop |
2 | IK101 | Ik104 | GFP loop |
PCR Polymerase : KAPA hifi
95.0°C : 5min
98.0°C : 20sec
58.0°C : 15sec
72.0°C : 40sec
72.0°C : 5min
4.0°C : Infinate
X35 cycle
10/15
PCR
yoshimoto
lane | Primer Foward | Primer Revese | Template |
---|
1 | IK85 | IK86 | Rev parts |
2 | IK87 | IK88 | Rev parts |
3 | IK61 | OD54 | T7-RRE-GFP |
PCR Polymerase : KAPA hifi
95.0°C : 5min
98.0°C : 20sec
58.0°C : 15sec
72.0°C : 40sec
72.0°C : 5min
4.0°C : Infinate
X30 cycle
10/17
PCR
yoshitomo
lane | Primer Foward | Primer Revese | Template |
---|
1 | IK61 | OD54 | 4 |
2 | IK61 | OD54 | 5 |
3 | IK61 | OD54 | 7 |
PCR Polymerase : KAPA hifi
95.0°C : 5min
98.0°C : 20sec
58.0°C : 15sec
72.0°C : 40sec
72.0°C : 5min
4.0°C : Infinate
X30 cycle
10/17
PCR
yoshimoto
lane | Primer Foward | Primer Revese | Template |
---|
1 | IK87 | IK88 | Rev |
2 | IK105 | IK108 | Gal1-loop |
PCR Polymerase :
95.0°C : 5min
98.0°C : 20sec
55.0°C : 15sec
72.0°C : 40sec
72.0°C : 5min
4.0°C : Infinate
X30 cycle
10/18
PCR
yoshimoto
lane | Primer Foward | Primer Revese | Template |
---|
1 | IK105 | IK108 | template1 |
PCR Polymerase :
95.0°C : 5min
98.0°C : 20sec
58.0°C : 15sec
72.0°C : 40sec
72.0°C : 5min
4.0°C : Infinate
X30 cycle
10/18
PCR
yoshimoto
lane | Primer Foward | Primer Revese | Template |
---|
1 | IK107 | IK108 | Gal1pro-loop |
2 | IK105 | IK106 | Gal1pro-loop |
PCR Polymerase :
95.0°C : 5min
98.0°C : 20sec
58.0°C : 15sec
72.0°C : 40sec
72.0°C : 5min
4.0°C : Infinate
X30 cycle
10/19
PCR
yoshimoto
lane | Primer Foward | Primer Revese | Template |
---|
1 | OD22 | OD23 | GPD-loop |
2 | IK87 | IK88 | Gal1-loop |
3 | IK105 | IK108 | T7-RRE |
4 | IK101 | IK104 | TDH3-eGFP |
5 | OD31 | OD32 | Rev |
PCR Polymerase :
95.0°C : 5min
98.0°C : 20sec
58.0°C : 15sec
72.0°C : 40sec
72.0°C : 5min
4.0°C : Infinate
X30 cycle
10/20
PCR
yoshimoto
lane | Primer Foward | Primer Revese | Template |
---|
1 | M13F | M13R | Colony 1-10 (RRE) |
2 | M13F | M13R | Colony11-17 (TDH3-eGFP) |
3 | M13F | M13R | Colony 18-21 (Rev) |
4 | M13F | M13R | Colony 22-23 (GPDp-loop) |
5 | M13F | M13R | Colony24-28 (GPDp-loop) |
6 | M13F | M13R | Colony 29-32 (T7-RRE) |
7 | M13F | M13R | Colony 33-35 (TDH3-eGFP) |
8 | M13F | M13R | Colony 36-37 (GPDp-loop) |
9 | M13F | M13R | Colony 38-40 (Gal1-loop) |
10 | M13F | M13R | Colony 41-43 (Rev) |
11 | M13F | M13R | Colony 44 (GPDp-loop) |
PCR Polymerase :
95.0°C : 5min
98.0°C : 20sec
58.0°C : 15sec
72.0°C : 40sec
72.0°C : 5min
4.0°C : Infinate
X30 cycle
10/22
PCR
yoshimoto
lane | Primer Foward | Primer Revese | Template |
---|
1 | OD22 | OD23 | TDH3 GFP |
2 | IK87 | IK88 | Rev |
3 | IK105 | IK108 | Gal1 |
4 | IK101 | IK104 | GPD1 |
5 | OD31 | OD32 | T7-RRE |
PCR Polymerase :
95.0°C : 5min
98.0°C : 20sec
58.0°C : 15sec
72.0°C : 40sec
72.0°C : 5min
4.0°C : Infinate
X30 cycle