Sensing part
pmrC-GFP→pmrC-GFP-pBAD30
7.10
PCR
Yiyan Yu
7.19
Restriction Enzyme Digestion
Yiyan Yu
Name |
Enzyme |
pmrC-GFP |
EcoRI and Hind III |
pBAD30 |
EcoRI and Hind III |
7.20
DNA ligation & transformation
Yiyan Yu & Qingyang Yu
Name |
Backbone |
Strain |
Medium |
pmrC-GFP |
pBAD30(EcoR I and Hind III cut) |
Top10 |
LB(amp added) |
7.21
Colony PCR
Yiyan Yu
Number |
Name |
1 |
pmrC-GFP-pBAD30 |
5 |
pmrC-GFP-pBAD30 |
6 |
pmrC-GFP-pBAD30 |
PmrA-PmrB(Fe)→PmrA-PmrB(Fe)-pBAD30
7.8
PCR
Long Cheng
Name |
Total |
PmrA-PmrB(Fe) |
50 μl |
7.19
Restriction Enzyme Digestion
Yiyan Yu
Name |
Enzyme |
PmrA-PmrB(Fe) |
EcoRI and KpnI |
pBAD30 |
EcoRI and KpnI |
7.20
Restriction Enzyme Digestion
Yiyan Yu & Qingyang Yu
Name |
Backbone |
Strain |
Medium |
PmrA-PmrB(Fe) |
pBAD30(EcoR I and Hind III cut) |
Top10 |
LB(amp added) |
7.21
Colony PCR
Yiyan Yu
Number |
Name |
1 |
PmrA-PmrB(Fe)-pBAD30 |
5 |
PmrA-PmrB(Fe)-pBAD30 |
6 |
PmrA-PmrB(Fe)-pBAD30 |
PmrA-PmrB(Fe)-pBAD30→PmrA-PmrB(LBTX)-pBAD30
7.25
PCR
Yiyan Yu
Name |
Total |
LBT 1 to 12 |
50 μl each |
8.10
Point mutation
Xiaoyang Li
Name |
Total |
PmrA-PmrB(Fe)-pBAD30 |
50 μl |
8.20
Restriction Enzyme Digestion
Xiaoyang Li
Name |
Enzyme |
PmrA-PmrB(Fe)-pBAD30 |
AhdI and SacI |
LBT 1 |
AhdI and SacI |
LBT 11 |
AhdI and SacI |
8.20
Transformation
Xiaoyang Li & Yating Hu
Name |
Backbone |
Strain |
Medium |
LBT 1 |
PmrA-PmrB(Fe)-pBAD30(AhdI and SacI cut) |
Top10 |
LB(amp added) |
LBT 11 |
PmrA-PmrB(Fe)-pBAD30(AhdI and SacI cut) |
Top10 |
LB(amp added) |
8.21
Colony PCR
Xiaoyang Li
Number |
Name |
1 |
PmrA-PmrB(LBT 1)-pBAD30 |
1 |
PmrA-PmrB(LBT 11)-pBAD30 |
8.31
Restriction Enzyme Digestion
Xiaoyang Li
Name |
Enzyme |
PmrA-PmrB(LBT2) |
EcoRI and KpnI |
PmrA-PmrB(LBT3) |
EcoRI and KpnI |
PmrA-PmrB(LBT4) |
EcoRI and KpnI |
PmrA-PmrB(LBT5) |
EcoRI and KpnI |
PmrA-PmrB(LBT6) |
EcoRI and KpnI |
PmrA-PmrB(LBT7) |
EcoRI and KpnI |
PmrA-PmrB(LBT8) |
EcoRI and KpnI |
PmrA-PmrB(LBT9) |
EcoRI and KpnI |
PmrA-PmrB(LBT10) |
EcoRI and KpnI |
PmrA-PmrB(LBT12) |
EcoRI and KpnI |
8.31
Transformation
Xiaoyang Li & Yating Hu
Name |
Backbone |
Strain |
Medium |
PmrA-PmrB(LBT 2,3,4,5,6,7,8,9,10,12) |
pBAD30(EcoR I and Kpn I cut) |
Top10 |
LB(amp added) |
9.3
Colony PCR
Xiaoyang Li
Number |
Name |
1 |
PmrA-PmrB(LBT2) |
2 |
PmrA-PmrB(LBT3) |
3 |
PmrA-PmrB(LBT4) |
4 |
PmrA-PmrB(LBT5) |
5 |
PmrA-PmrB(LBT6) |
6 |
PmrA-PmrB(LBT7) |
7 |
PmrA-PmrB(LBT8) |
8 |
PmrA-PmrB(LBT9) |
9 |
PmrA-PmrB(LBT10) |
10 |
PmrA-PmrB(LBT12) |
PmrA-PmrB(LBT X)-pBAD30→PmrA-PmrB(LBT X)-pmrC-GFP-pBAD30
10.18
PCR
Long Cheng
Name |
Total |
PmrA-PmrB(LBT 5) |
30 μl |
PmrA-PmrB(LBT 9) |
30 μl |
PmrA-PmrB(LBT 11) |
30 μl |
PmrC-GFP |
50 μl |
10.19
Bypass PCR
Xiaoyang Li & Qingyang Yu
Name |
Total |
PmrA-PmrB(LBT 5) & PmrC-GFP |
50 μl |
PmrA-PmrB(LBT 9) & PmrC-GFP |
50 μl |
PmrA-PmrB(LBT 11) & PmrC-GFP |
50 μl |
10.19
Restriction Enzyme Digestion
Xiaoyang Li
Name |
Enzyme |
PmrA-PmrB(LBT 5) & PmrC-GFP |
EcoRI and Hind III |
PmrA-PmrB(LBT 9) & PmrC-GFP |
EcoRI and Hind III |
PmrA-PmrB(LBT 11) & PmrC-GFP |
EcoRI and Hind III |
pBAD30 |
EcoRI and Hind III |
10.19
DNA ligation & transformation
Xiaoyang Li & Yating Hu
Name |
Backbone |
Strain |
Medium |
PmrA-PmrB(LBT 5)-PmrC-GFP |
pBAD30(EcoRI and Hind III cut) |
Top10 |
LB(amp added) |
PmrA-PmrB(LBT 9)-PmrC-GFP |
pBAD30(EcoRI and Hind III cut) |
Top10 |
LB(amp added) |
PmrA-PmrB(LBT 11)-PmrC-GFP |
pBAD30(EcoRI and Hind III cut) |
Top10 |
LB(amp added) |
10.20
Colony PCR
Xiaoyang Li
Number |
Name |
1 |
PmrA-PmrB(LBT 5)-PmrC-GFP |
1 |
PmrA-PmrB(LBT 9)-PmrC-GFP |
1 |
PmrA-PmrB(LBT 11)-PmrC-GFP |
Standardization Part
pmrA-pmrB-LBT X-pBAD30→pmrA-pmrB-LBT X-pmrC-GFP-pBAD30
8.20
PCR
Yubei Sun & Huiping Shi
Name |
Total |
oprF-3 X LBT1 |
50 μl |
oprF-3 X LBT4 |
50 μl |
oprF-3 X LBT8 |
50 μl |
oprF-3 X LBT10 |
50 μl |
oprF-3 X LBT11 |
50 μl |
oprF-3 X LBT12 |
50 μl |
9.20
Bypass PCR
Efan Wang
Name |
Total |
oprF-3 X LBT3 |
50 μl |
oprF-3 X LBT6 |
50 μl |
oprF-3 X LBT7 |
50 μl |
oprF-Sitag |
50 μl |
9.5
Restriction Enzyme Digestion
Yubei Sun & Ziyi Li
Name |
Enzyme |
oprF-3 X LBT 1 |
EcoRI and Spel I |
oprF-3 X LBT 4 |
EcoRI and Spel I |
oprF-3 X LBT 10 |
EcoRI and Spel I |
oprF-3 X LBT 11 |
EcoRI and Spel I |
pSB1C3 |
EcoRI and Spel I |
pSB1C3 |
Xbal I and Pst I |
9.20
Restriction Enzyme Digestion
Huiping Shi
Name |
Enzyme |
oprF-3 X LBT 3 |
EcoRI and Spel I |
oprF-3 X LBT 6 |
EcoRI and Spel I |
oprF-3 X LBT 7 |
EcoRI and Spel I |
oprF-3 X LBT 8 |
EcoRI and Spel I |
oprF-3 X LBT 11 |
EcoRI and Spel I |
oprF-3 X LBT 12 |
EcoRI and Spel I |
9.27
DNA ligation & transformation
Yubei Sun & Huiping Shi
Name |
Backbone |
Strain |
Medium |
oprF-3 X LBT 1 |
pSB1C3(EcoRI and Spel I cut) |
DH5a |
LB(chl added) |
oprF-3 X LBT 3 |
pSB1C3(EcoRI and Spel I cut) |
DH5a |
LB(chl added) |
oprF-3 X LBT 4 |
pSB1C3(EcoRI and Spel I cut) |
DH5a |
LB(chl added) |
oprF-3 X LBT 8 |
pSB1C3(EcoRI and Spel I cut) |
DH5a |
LB(chl added) |
oprF-Sitag |
pSB1C3(EcoRI and Spel I cut) |
DH5a |
LB(chl added) |
9.27
DNA ligation & transformation
Ziyang Xiao
Name |
Backbone |
Strain |
Medium |
oprF-3 X LBT 6 |
pSB1C3(EcoRI and Spel I cut) |
DH5a |
LB(chl added) |
oprF-3 X LBT 7 |
pSB1C3(EcoRI and Spel I cut) |
DH5a |
LB(chl added) |
oprF-3 X LBT 10 |
pSB1C3(EcoRI and Spel I cut) |
DH5a |
LB(chl added) |
oprF-3 X LBT 11 |
pSB1C3(EcoRI and Spel I cut) |
DH5a |
LB(chl added) |
oprF-3 X LBT 12 |
pSB1C3(EcoRI and Spel I cut) |
DH5a |
LB(chl added) |
9.28
Colony PCR
Yubei Sun
Number |
Name |
1 |
oprF-3×LBT1-pSB1C3 |
2 |
oprF-3×LBT3-pSB1C3 |
3 |
oprF-3×LBT4-pSB1C3 |
4 |
oprF-3×LBT8-pSB1C3 |
5 |
oprF-Sitag-pSB1C3 |
9.30
Colony PCR
Ziyang Xiao
Number |
Name |
1 |
oprF-3×LBT6-pSB1C3 |
2 |
oprF-3×LBT7-pSB1C3 |
3 |
oprF-3×LBT10-pSB1C3 |
4 |
oprF-3×LBT11-pSB1C3 |
5 |
oprF-3×LBT12-pSB1C3 |
pmrC-GFP-pBAD30→pmrC-GFP-pSB1C3
10.2
PCR
Ziyang Xiao
Name |
Total |
pmrC-GFP |
30 μl |
10.4
Restriction Enzyme Digestion
Yubei Sun
Name |
Enzyme |
pmrC-GFP |
Pstl I and XbaI |
10.5
DNA ligation & transformation
Ziyang Xiao
Name |
Backbone |
Strain |
Medium |
pmrC-GFP |
pSB1C3(Pstl I and XbaI cut) |
DH5a |
LB(chl added) |
10.6
Colony PCR
Ziyang Xiao
Number |
Name |
1 |
pmrC-GFP-pSB1C3 |
2 |
pmrC-GFP-pSB1C3 |
3 |
pmrC-GFP-pSB1C3 |
4 |
pmrC-GFP-pSB1C3 |
5 |
pmrC-GFP-pSB1C3 |
pmrA-pmrB-LBT X-pmrC-GFP-pBAD30→pmrA-pmrB-LBT X-pmrC-GFP-pSB1C3
10.26
PCR
Ziyang Xiao
Name |
Total |
pmrA-pmrB-LBT 5-pmrC-GFP |
30 μl |
pmrA-pmrB-LBT 9-pmrC-GFP |
30 μl |
pmrA-pmrB-LBT 11-pmrC-GFP |
30 μl |
10.4
Restriction Enzyme Digestion
Huiping Shi
Name |
Enzyme |
pmrA-pmrB-LBT 5-pmrC-GFP |
Pst I and XbaI |
pmrA-pmrB-LBT 9-pmrC-GFP |
Pst I and XbaI |
pmrA-pmrB-LBT 11-pmrC-GFP |
Pst I and XbaI |
10.5
DNA ligation & transformation
Huiping Shi
Name |
Backbone |
Strain |
Medium |
pmrA-pmrB-LBT 5-pmrC-GFP |
pSB1C3(Pst I and Spel I cut) |
DH5a |
LB(chl added) |
pmrA-pmrB-LBT 9-pmrC-GFP |
pSB1C3(Pst I and Spel I cut) |
DH5a |
LB(chl added) |
pmrA-pmrB-LBT 11-pmrC-GFP |
pSB1C3(Pst I and Spel I cut) |
DH5a |
LB(chl added) |
10.6
Colony PCR
Ziyang Xiao
Number |
Name |
1 |
pmrA-pmrB-LBT 5-pmrC-GFP |
2 |
pmrA-pmrB-LBT 9-pmrC-GFP |
3 |
pmrA-pmrB-LBT 11-pmrC-GFP |