|
|
Line 180: |
Line 180: |
| | | |
| </ul> | | </ul> |
− | </div> | + | <p>7.23 |
| + | 1、PCR for BioBricks, 5’ homologous arm, backbone |
| + | 2、electrophoresis:Validate PCR product |
| + | 3、枯草杆菌保种 |
| | | |
− | <div class="column full_size">
| + | 7.24 |
| + | 1、Get genome |
| + | 2、昨日PCR产物回收 |
| + | 3、Genome PCR |
| + | 5、引物退火 |
| + | 6、transcribe, replicate pVeg |
| | | |
− | <h1>Notebook</h1>
| + | 7.26 |
− | <p> Document the dates you worked on your project. This should be a detailed account of the work done each day for your project.</p>
| + | 1、electrophoresis(genome,退火产物) |
| | | |
− | </div>
| + | 8.3 |
− | <div class="clear"></div>
| + | 1、enzyme digestion: pHT01, using BamHI & XbaI |
| + | 2、electrophoresis: testing the product of enzyme digestion(PHT01,PHT01 enzyme digestion product,pVeg PCR, pVeg PCR enzyme digestion product) |
| + | 10、胶回收 |
| | | |
| + | 8.4 |
| + | 1、PCR(pVeg,GFP, negative control,positive control) |
| + | 2、 electrophoresis: test the product of PCR |
| + | 5、enzyme digestion: YerP, gfp, sfp, pSB1C3 |
| + | 6、Ligation |
| + | 7、PCR产物回收 |
| + | 8、transcription |
| | | |
− | <div class="column half_size">
| + | 8.5 |
− | <h5>What should this page have?</h5>
| + | 1、enzyme digestion: pVeg, RBS, Backbone |
− | <ul>
| + | 2、Colony PCR(sfp2 A 抗, YerP1 A抗, YerP2 A抗) |
− | <li>Chronological notes of what your team is doing.</li>
| + | |
− | <li> Brief descriptions of daily important events.</li>
| + | |
− | <li>Pictures of your progress. </li>
| + | |
− | <li>Mention who participated in what task.</li>
| + | |
− | </ul>
| + | |
| | | |
− | </div>
| + | 8.9 |
| + | 1、electrophoresis: 菌PCR |
| + | 2、get genome,ER,尝试双酶切验证质粒 |
| | | |
− | <div class="column half_size">
| + | 8.10 |
− | <h5>Inspiration</h5>
| + | 1、B.sub 感受态制备 |
− | <p>You can see what others teams have done to organize their notes:</p>
| + | 2、8月9号转化涂板因未吹干失败,菌落被感染 |
| | | |
− | <ul>
| + | 8.14 |
− | <li><a href="https://2014.igem.org/Team:ATOMS-Turkiye/Notebook">2014 ATOMS-Turkiye</a></li>
| + | 1、enzyme digestion: sfp,pSB1C3 |
− | <li><a href="https://2014.igem.org/Team:Tec-Monterrey/ITESM14_project.html#tab_notebook">2014 Tec Monterrey</a></li>
| + | 2、electrophoresis: test the product of胶纯化 |
− | <li><a href="https://2014.igem.org/Team:Kyoto/Notebook/Magnetosome_Formation#title">2014 Kyoto</a></li>
| + | 3、transcribe RFP Construct(control) |
− | <li><a href="https://2014.igem.org/Team:Cornell/notebook">2014 Cornell</a></li>
| + | 4、ligate 19-TC 载体,product(YerP /sfp),Solution Ⅰ Control |
− | </ul>
| + | |
| | | |
− | </div>
| + | 8.21 |
| + | 1、PCR(sfp,YerP,pVeg,positive control,negative control) |
| + | 3、ligation pVeg-RBS A抗 |
| | | |
| + | 8.21 |
| + | 1、Transformation(pVeg DH5α C抗,pVeg RBF DH5α A抗,D6B HP K抗,PUC19 对照A抗(稀),rfp枯草未知1,rfp枯草未知2,PUC19对照A抗(浓), pVeg DH5α A抗,rfp DH5α K抗,pVeg RBS A抗,sfp, YerP) |
| + | |
| + | 8.22 |
| + | 1、Taq Super-mix |
| + | 2、electrophoresis |
| + | 3、Digestion |
| + | 4、Ligation |
| + | |
| + | 8.23 |
| + | 1、electrophoresis(D2000,1kb Plus Ladder,pSB1A3-GFP,pSB1C3-pTet-GFP,positive control,negative control) |
| + | 2、digestion for validation |
| + | 3、electrophoresis(ladder, pveg,pveg切,pveg-RBS,pveg-RBS 切,pH101-RFP,pH101-RFP切,T-sfp,T-sfp 旧,T-YerP,sfp-RBS) |
| + | |
| + | 9.2 |
| + | 1、PCR plasmid(T-sfp提,T-YerP提,sfp-RBS提,pvg-RBS提,T-sfp连,T-YerP连,sfp-RBS连,pvg-RBS连,pveg-RBS) |
| + | 2、electrophoresis(ladder,pSB1A3-GFPpSB1C3-pTet-GFP丙sfp2 A 抗, 庚, YerP1 A抗, 辛, negative control,positive control)(ladder,TSFP1,TSFP2,TSFP3,TSFP4,TSFP5) |
| + | (ladder,T-SFP,T-SFP切,sfp-RBS,sfp-RBS切,pVg-RBS,pVg-RBS切,T-YerP,T-YerP切,PHT01-GFP,PHT01-GFP切)(ladder,pSB1A3-GFP,己,T-YerP1,T-YerP2,T-YerP3,T-YerP4,T-YerP5,T-YerP6,T-YerP7) |
| + | |
| + | 第19页左半部分不知道说什么 |
| + | |
| + | 第20页最上面有个5号可能是电泳 |
| + | |
| + | 9.4 |
| + | 1、transcribe T-sfp,T-YerP |
| + | |
| + | 9.5 |
| + | 1、PCR(5‘Homologous arm,3’Homologous arm,backbone,BioBricks site) |
| + | 2、胶回收 |
| + | 3、Transcribe(T-sfp,Y-YerP,pVeg,positive control) |
| + | |
| + | 9.8 |
| + | 1、PCR |
| + | 2、PCR:T-sfp |
| + | 3、enzyme digestion(E、P),(PSB1C3-rfp plasmid) |
| + | 4、胶回收 |
| + | 5、验证(ladder,5‘homologous arm,3‘homologous arm,bb,RFP construct) |
| + | |
| + | 9.9 |
| + | 1、electrophoresis: test pSB1C3-rfp回收plasmid及7.26PCR得YerP |
| + | 2、Pfu扩增Biobrick sites及k位点 |
| + | 3、上样(1-6 6管PSB1C3-rfp, 7-8 7.26 YerP,PSB1A3-gfp质粒) |
| + | 4、YerP平板取菌划线,菌落PCR |
| + | 7、get plasmid→PSB1C3-rfp construct,PCR |
| + | 9、electrophoresis: test product of PSB1C3-rfp 回收产物 |
| + | 10、凝胶回收(sfp-biobrick, pVeg) |
| + | |
| + | 9.9 |
| + | 1、胶回收 |
| + | 2、electrophoresis1(D2000, RFP1,RFP2,RFP3,Bsub2,Bsub3,Bsub4,Busb5,Bsub6) |
| + | 3、electrophoresis(D2000, Biobrick,sfp,Pveg) |
| + | |
| + | 9.12 |
| + | 1、enzyme digestion(PSB1C3-rfp) |
| + | 2、electrophoresis——胶回收 |
| + | (左:ladder,bs,5‘homologous arm,3‘homologous arm,BioBricks site,pSB1C3) |
| + | (右:ladder,YerP1,YerP2,YerP3,YerP4,pSB1C3) |
| + | 3、test(ladder,pVeg-RBS,sfp-RBS,sfp-bb,A3,A4,A5,A6) |
| + | |
| + | 9.13 |
| + | 1、胶回收 |
| + | 2、PCR(pVeg-RBS,sfp-RBS,ext2) |
| + | 3、7管PHT01酶切检测 |
| + | 4、第二轮RBS PCR体系 |
| + | 6、electrophoresis:胶回收产物回收(ladder,YerP1,YerP2,YerP3,YerP4,BioBricks site,Backbone,5‘ homologous arm,3‘ homologous arm,BioBricks site) |
| + | PCR产物 |
| + | 1、 ladder-D2000 |
| + | 2、 pVg-RBS-ext 1 |
| + | 3、 pVg-RBS-ext 2 |
| + | 4、 sfp-RBS-ext 1 |
| + | 5、 sfp-RBS-ext 2 |
| + | 6、 PSB1C3回收 |
| + | |
| + | 9.14 |
| + | 1、electrophoresis (ladder:1kb plus DNA) |
| + | 2、pVeg-RBS-ext2 |
| + | 3、sfp-RBS-ext2 |
| + | 4、pHT01 1 |
| + | 5、pHT01 2 |
| + | |
| + | 9.15 |
| + | 1、pfu: erP+BioBricks site PCR |
| + | 2、加A |
| + | 3、YerP 加A |
| + | 4、electrophoresis(1kb ladder,YerP1 bio,YerP2 bio) |
| + | 5、electrophoresis(ladder 1kb plus,pVeg-RBS 加A 回收,sfp-RBS 加A 回收,YerP1 回收,YerP2 回收,PSB1C-sfp,RFP3) |
| + | 6、连接 |
| + | |
| + | 9.17 |
| + | 1、transcribe: Bsub |
| + | 2、transcribe: T-YerP |
| + | 3、OEPCR1 |
| + | |
| + | 9.18 |
| + | 1、枯草感受态检测——菌落数量 |
| + | 2、回收pVeg+sfp OEPCR |
| + | 3、重叠PCR体系 |
| + | |
| + | 9.20 |
| + | 1、OEPCR3 |
| + | 2、菌落PCR(psb1C3-AH) |
| + | 3、菌PCR electrophoresis(ladder,YerP,sfp,GFP,negative control,positive control,6) |
| + | 4、回收1000kb左右的pvg-sfp |
| + | 5、回收OEPCR3 |
| + | |
| + | 9.21 |
| + | 1、新backbone,BioBricks site用pfu和2017○34GP3 |
| + | |
| + | 9.22 |
| + | 1、YerP PCR |
| + | 2、胶回收 |
| + | |
| + | 9.25 |
| + | 1、PCR:backbone,BioBricks site |
| + | |
| + | 9.26 |
| + | 1、OEPCR3 加A |
| + | 2、electrophoresis(none,ladder D2000,BioBricks site,backbone) |
| + | |
| + | 9.27 |
| + | 1、electrophoresis: for testing the product of回收产物 |
| + | First: 2-5道:T-YerP○5 1-4; 6-9道:T-YerP○4 1-4) |
| + | Second:(2-5道:AH,1-4;6-7道:BioBricks site,Backbone) |
| + | 2、transcribe pVeg-bfp |
| + | |
| + | 9.28 |
| + | 1、菌PCR(ladder D2000,T-YerP 10,T-YerP 11,T-YerP 9,T-YerP 5,T-YerP 4,T-YerP pvg-sfp,T-YerP粉,划线5,划线4) |
| + | |
| + | 9.30 |
| + | 1、菌液PCR |
| + | 2、Gibson片段-pveg-sfp PCR) |
| + | 3、electrophoresis(1,2,3,4,5,pveg sfp gibs,+,-) |
| + | |
| + | 10.9 |
| + | 1、回收Gib-(pveg-sfp) |
| + | 2、enzyme digestion pVeg-sfp |
| + | 3、ligate pSB1C3-pvg-sfp |
| + | |
| + | 10.10 |
| + | 1、electrophoresis(ladder,YerP1,YerP2,YerP3,YerP4,5‘ homologous arm,3‘ homologous arm,Gb,bb,BB) |
| + | |
| + | 10.11 |
| + | 1、electrophoresis(D2000,1,2,3,4,5,6,7,8,9,10) |
| + | 2、electrophoresis(D2000,11,12,13,14,15,16,17,18) |
| + | |
| + | 10.16 |
| + | 1、electrophoresis(ladder D2000,OEPCR3) |
| + | 2、transcribe(control,T-pveg sfp rbs,IDT’s ImrA) |
| + | 3、electrophoresis testing(1kb plus,OEPCR 回收产物,5‘,3‘,pSB1C3切, p SB1C3) |
| + | |
| + | 10.18 |
| + | 1、PCR菌(○1pSB1A3-GFPpSB1C3-pTet-GFP丙丁戊阴○2psb-pvg-sfp连接产物,OEPCR,RFP3,P3FG) |
| + | |
| + | 10.19 |
| + | 1、菌PCR(○1ladder 2000,sfp,GFP,negative control,positive control,6)(D2000,psb-pvg-sfp连接产物,OEPCR,RFP3,P3FG) |
| + | |
| + | 10.21 |
| + | 1、PCR |
| + | 2、enzyme digestion(psb1C3,RFP3,lmrA质粒,pvg-sfp) |
| + | 3、electrophoresis(ladder 1kb plus,OEPCR3,ImrA1,ImrA2,D1,E1) |
| + | 4、electrophoresis(ladder 1kb plus,A1,B1,C1) |
| + | |
| + | |
| + | 10.22 |
| + | 1、37度恒温箱里(lmrA切1,2(E,P得PCR产物),超薄试剂盒纯化,验证 |
| + | 2、Ligate PSB1C3-lmrA |
| + | 3、 Transcribe PSB1C3-pveg-sfp转化得平板 |
| + | 4、Gibson |
| + | 6、gov electrophoresis(1kb plus marker,lmrA切1,2回收,lmrA2*2,lmrA3*2,pvg sfp*2,PSB1C3线性,pSB1C3) |
| + | |
| + | |
| + | 3、菌PCR(D2000,浓pveg-sfp,OEPCR,阳) |
| + | 4、菌PCR(D2000,YerP,sfp) |
| + | |
| + | 10.23 |
| + | 1、electrophoresis(D2000, ImrA2,lmrA2,lmrA3,lmrA4) |
| + | |
| + | 10.24 |
| + | 1、electrophoresis(ladder 1kb plus,lmrA2,lmrA3,lmrA4,1-1 pvg-rbs,2-1 pvg-rbs,2-3 pvg-rbs,1-2 pvg-rbs,1-3 pvg-rbs,RFP |
| + | </p> |
| </body> | | </body> |
| | | |
| </html> | | </html> |
RDFZ-China
7.23
1、PCR for BioBricks, 5’ homologous arm, backbone
2、electrophoresis:Validate PCR product
3、枯草杆菌保种
7.24
1、Get genome
2、昨日PCR产物回收
3、Genome PCR
5、引物退火
6、transcribe, replicate pVeg
7.26
1、electrophoresis(genome,退火产物)
8.3
1、enzyme digestion: pHT01, using BamHI & XbaI
2、electrophoresis: testing the product of enzyme digestion(PHT01,PHT01 enzyme digestion product,pVeg PCR, pVeg PCR enzyme digestion product)
10、胶回收
8.4
1、PCR(pVeg,GFP, negative control,positive control)
2、 electrophoresis: test the product of PCR
5、enzyme digestion: YerP, gfp, sfp, pSB1C3
6、Ligation
7、PCR产物回收
8、transcription
8.5
1、enzyme digestion: pVeg, RBS, Backbone
2、Colony PCR(sfp2 A 抗, YerP1 A抗, YerP2 A抗)
8.9
1、electrophoresis: 菌PCR
2、get genome,ER,尝试双酶切验证质粒
8.10
1、B.sub 感受态制备
2、8月9号转化涂板因未吹干失败,菌落被感染
8.14
1、enzyme digestion: sfp,pSB1C3
2、electrophoresis: test the product of胶纯化
3、transcribe RFP Construct(control)
4、ligate 19-TC 载体,product(YerP /sfp),Solution Ⅰ Control
8.21
1、PCR(sfp,YerP,pVeg,positive control,negative control)
3、ligation pVeg-RBS A抗
8.21
1、Transformation(pVeg DH5α C抗,pVeg RBF DH5α A抗,D6B HP K抗,PUC19 对照A抗(稀),rfp枯草未知1,rfp枯草未知2,PUC19对照A抗(浓), pVeg DH5α A抗,rfp DH5α K抗,pVeg RBS A抗,sfp, YerP)
8.22
1、Taq Super-mix
2、electrophoresis
3、Digestion
4、Ligation
8.23
1、electrophoresis(D2000,1kb Plus Ladder,pSB1A3-GFP,pSB1C3-pTet-GFP,positive control,negative control)
2、digestion for validation
3、electrophoresis(ladder, pveg,pveg切,pveg-RBS,pveg-RBS 切,pH101-RFP,pH101-RFP切,T-sfp,T-sfp 旧,T-YerP,sfp-RBS)
9.2
1、PCR plasmid(T-sfp提,T-YerP提,sfp-RBS提,pvg-RBS提,T-sfp连,T-YerP连,sfp-RBS连,pvg-RBS连,pveg-RBS)
2、electrophoresis(ladder,pSB1A3-GFPpSB1C3-pTet-GFP丙sfp2 A 抗, 庚, YerP1 A抗, 辛, negative control,positive control)(ladder,TSFP1,TSFP2,TSFP3,TSFP4,TSFP5)
(ladder,T-SFP,T-SFP切,sfp-RBS,sfp-RBS切,pVg-RBS,pVg-RBS切,T-YerP,T-YerP切,PHT01-GFP,PHT01-GFP切)(ladder,pSB1A3-GFP,己,T-YerP1,T-YerP2,T-YerP3,T-YerP4,T-YerP5,T-YerP6,T-YerP7)
第19页左半部分不知道说什么
第20页最上面有个5号可能是电泳
9.4
1、transcribe T-sfp,T-YerP
9.5
1、PCR(5‘Homologous arm,3’Homologous arm,backbone,BioBricks site)
2、胶回收
3、Transcribe(T-sfp,Y-YerP,pVeg,positive control)
9.8
1、PCR
2、PCR:T-sfp
3、enzyme digestion(E、P),(PSB1C3-rfp plasmid)
4、胶回收
5、验证(ladder,5‘homologous arm,3‘homologous arm,bb,RFP construct)
9.9
1、electrophoresis: test pSB1C3-rfp回收plasmid及7.26PCR得YerP
2、Pfu扩增Biobrick sites及k位点
3、上样(1-6 6管PSB1C3-rfp, 7-8 7.26 YerP,PSB1A3-gfp质粒)
4、YerP平板取菌划线,菌落PCR
7、get plasmid→PSB1C3-rfp construct,PCR
9、electrophoresis: test product of PSB1C3-rfp 回收产物
10、凝胶回收(sfp-biobrick, pVeg)
9.9
1、胶回收
2、electrophoresis1(D2000, RFP1,RFP2,RFP3,Bsub2,Bsub3,Bsub4,Busb5,Bsub6)
3、electrophoresis(D2000, Biobrick,sfp,Pveg)
9.12
1、enzyme digestion(PSB1C3-rfp)
2、electrophoresis——胶回收
(左:ladder,bs,5‘homologous arm,3‘homologous arm,BioBricks site,pSB1C3)
(右:ladder,YerP1,YerP2,YerP3,YerP4,pSB1C3)
3、test(ladder,pVeg-RBS,sfp-RBS,sfp-bb,A3,A4,A5,A6)
9.13
1、胶回收
2、PCR(pVeg-RBS,sfp-RBS,ext2)
3、7管PHT01酶切检测
4、第二轮RBS PCR体系
6、electrophoresis:胶回收产物回收(ladder,YerP1,YerP2,YerP3,YerP4,BioBricks site,Backbone,5‘ homologous arm,3‘ homologous arm,BioBricks site)
PCR产物
1、 ladder-D2000
2、 pVg-RBS-ext 1
3、 pVg-RBS-ext 2
4、 sfp-RBS-ext 1
5、 sfp-RBS-ext 2
6、 PSB1C3回收
9.14
1、electrophoresis (ladder:1kb plus DNA)
2、pVeg-RBS-ext2
3、sfp-RBS-ext2
4、pHT01 1
5、pHT01 2
9.15
1、pfu: erP+BioBricks site PCR
2、加A
3、YerP 加A
4、electrophoresis(1kb ladder,YerP1 bio,YerP2 bio)
5、electrophoresis(ladder 1kb plus,pVeg-RBS 加A 回收,sfp-RBS 加A 回收,YerP1 回收,YerP2 回收,PSB1C-sfp,RFP3)
6、连接
9.17
1、transcribe: Bsub
2、transcribe: T-YerP
3、OEPCR1
9.18
1、枯草感受态检测——菌落数量
2、回收pVeg+sfp OEPCR
3、重叠PCR体系
9.20
1、OEPCR3
2、菌落PCR(psb1C3-AH)
3、菌PCR electrophoresis(ladder,YerP,sfp,GFP,negative control,positive control,6)
4、回收1000kb左右的pvg-sfp
5、回收OEPCR3
9.21
1、新backbone,BioBricks site用pfu和2017○34GP3
9.22
1、YerP PCR
2、胶回收
9.25
1、PCR:backbone,BioBricks site
9.26
1、OEPCR3 加A
2、electrophoresis(none,ladder D2000,BioBricks site,backbone)
9.27
1、electrophoresis: for testing the product of回收产物
First: 2-5道:T-YerP○5 1-4; 6-9道:T-YerP○4 1-4)
Second:(2-5道:AH,1-4;6-7道:BioBricks site,Backbone)
2、transcribe pVeg-bfp
9.28
1、菌PCR(ladder D2000,T-YerP 10,T-YerP 11,T-YerP 9,T-YerP 5,T-YerP 4,T-YerP pvg-sfp,T-YerP粉,划线5,划线4)
9.30
1、菌液PCR
2、Gibson片段-pveg-sfp PCR)
3、electrophoresis(1,2,3,4,5,pveg sfp gibs,+,-)
10.9
1、回收Gib-(pveg-sfp)
2、enzyme digestion pVeg-sfp
3、ligate pSB1C3-pvg-sfp
10.10
1、electrophoresis(ladder,YerP1,YerP2,YerP3,YerP4,5‘ homologous arm,3‘ homologous arm,Gb,bb,BB)
10.11
1、electrophoresis(D2000,1,2,3,4,5,6,7,8,9,10)
2、electrophoresis(D2000,11,12,13,14,15,16,17,18)
10.16
1、electrophoresis(ladder D2000,OEPCR3)
2、transcribe(control,T-pveg sfp rbs,IDT’s ImrA)
3、electrophoresis testing(1kb plus,OEPCR 回收产物,5‘,3‘,pSB1C3切, p SB1C3)
10.18
1、PCR菌(○1pSB1A3-GFPpSB1C3-pTet-GFP丙丁戊阴○2psb-pvg-sfp连接产物,OEPCR,RFP3,P3FG)
10.19
1、菌PCR(○1ladder 2000,sfp,GFP,negative control,positive control,6)(D2000,psb-pvg-sfp连接产物,OEPCR,RFP3,P3FG)
10.21
1、PCR
2、enzyme digestion(psb1C3,RFP3,lmrA质粒,pvg-sfp)
3、electrophoresis(ladder 1kb plus,OEPCR3,ImrA1,ImrA2,D1,E1)
4、electrophoresis(ladder 1kb plus,A1,B1,C1)
10.22
1、37度恒温箱里(lmrA切1,2(E,P得PCR产物),超薄试剂盒纯化,验证
2、Ligate PSB1C3-lmrA
3、 Transcribe PSB1C3-pveg-sfp转化得平板
4、Gibson
6、gov electrophoresis(1kb plus marker,lmrA切1,2回收,lmrA2*2,lmrA3*2,pvg sfp*2,PSB1C3线性,pSB1C3)
3、菌PCR(D2000,浓pveg-sfp,OEPCR,阳)
4、菌PCR(D2000,YerP,sfp)
10.23
1、electrophoresis(D2000, ImrA2,lmrA2,lmrA3,lmrA4)
10.24
1、electrophoresis(ladder 1kb plus,lmrA2,lmrA3,lmrA4,1-1 pvg-rbs,2-1 pvg-rbs,2-3 pvg-rbs,1-2 pvg-rbs,1-3 pvg-rbs,RFP