Difference between revisions of "Team:RDFZ-China/Notebook"

 
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<p>7.23
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1、PCR for BioBricks, 5’ homologous arm, backbone
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2、electrophoresis:Validate PCR product
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3、枯草杆菌保种
  
<div class="column full_size">
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7.24
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1、Get genome
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2、昨日PCR产物回收
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3、Genome PCR
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5、引物退火
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6、transcribe, replicate pVeg
  
<h1>Notebook</h1>
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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>
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1、electrophoresis(genome,退火产物)
  
</div>
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8.3
<div class="clear"></div>
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1、enzyme digestion: pHT01, using BamHI & XbaI
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2、electrophoresis: testing the product of enzyme digestion(PHT01,PHT01 enzyme digestion product,pVeg PCR, pVeg PCR enzyme digestion product)
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10、胶回收
  
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8.4
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1、PCR(pVeg,GFP, negative control,positive control)
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2、 electrophoresis: test the product of PCR
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5、enzyme digestion: YerP, gfp, sfp, pSB1C3
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6、Ligation
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7、PCR产物回收
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8、transcription
  
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8.5
<h5>What should this page have?</h5>
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1、enzyme digestion: pVeg, RBS, Backbone
<ul>
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2、Colony PCR(sfp2 A 抗, YerP1 A抗, YerP2 A抗)
<li>Chronological notes of what your team is doing.</li>
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<li> Brief descriptions of daily important events.</li>
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<li>Pictures of your progress. </li>
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<li>Mention who participated in what task.</li>
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</ul>
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</div>
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8.9
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1、electrophoresis: 菌PCR
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2、get genome,ER,尝试双酶切验证质粒
  
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8.10
<h5>Inspiration</h5>
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1、B.sub 感受态制备
<p>You can see what others teams have done to organize their notes:</p>
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2、8月9号转化涂板因未吹干失败,菌落被感染
  
<ul>
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8.14
<li><a href="https://2014.igem.org/Team:ATOMS-Turkiye/Notebook">2014 ATOMS-Turkiye</a></li>
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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>
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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>
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3、transcribe RFP Construct(control)
<li><a href="https://2014.igem.org/Team:Cornell/notebook">2014 Cornell</a></li>
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4、ligate 19-TC 载体,product(YerP /sfp),Solution Ⅰ Control
</ul>
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</div>
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8.21
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1、PCR(sfp,YerP,pVeg,positive control,negative control)
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3、ligation pVeg-RBS A抗
  
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8.21
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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)
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8.22
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1、Taq Super-mix
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2、electrophoresis
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3、Digestion
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4、Ligation
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8.23
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1、electrophoresis(D2000,1kb Plus Ladder,pSB1A3-GFP,pSB1C3-pTet-GFP,positive control,negative control)
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2、digestion for validation
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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)
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 +
9.2
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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)
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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)
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(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)
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第19页左半部分不知道说什么
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第20页最上面有个5号可能是电泳
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9.4
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1、transcribe T-sfp,T-YerP
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9.5
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1、PCR(5‘Homologous arm,3’Homologous arm,backbone,BioBricks site)
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2、胶回收
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3、Transcribe(T-sfp,Y-YerP,pVeg,positive control)
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9.8
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1、PCR
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2、PCR:T-sfp
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3、enzyme digestion(E、P),(PSB1C3-rfp plasmid)
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4、胶回收
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5、验证(ladder,5‘homologous arm,3‘homologous arm,bb,RFP construct)
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9.9
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1、electrophoresis: test pSB1C3-rfp回收plasmid及7.26PCR得YerP
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2、Pfu扩增Biobrick sites及k位点
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3、上样(1-6 6管PSB1C3-rfp, 7-8 7.26 YerP,PSB1A3-gfp质粒)
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4、YerP平板取菌划线,菌落PCR
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7、get plasmid→PSB1C3-rfp construct,PCR
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9、electrophoresis: test product of PSB1C3-rfp 回收产物
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10、凝胶回收(sfp-biobrick, pVeg)
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9.9
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1、胶回收
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2、electrophoresis1(D2000, RFP1,RFP2,RFP3,Bsub2,Bsub3,Bsub4,Busb5,Bsub6)
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3、electrophoresis(D2000, Biobrick,sfp,Pveg)
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9.12
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1、enzyme digestion(PSB1C3-rfp)
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2、electrophoresis——胶回收
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(左:ladder,bs,5‘homologous arm,3‘homologous arm,BioBricks site,pSB1C3)
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(右:ladder,YerP1,YerP2,YerP3,YerP4,pSB1C3)
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3、test(ladder,pVeg-RBS,sfp-RBS,sfp-bb,A3,A4,A5,A6)
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9.13
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1、胶回收
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2、PCR(pVeg-RBS,sfp-RBS,ext2)
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3、7管PHT01酶切检测
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4、第二轮RBS PCR体系
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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)
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PCR产物
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1、 ladder-D2000
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2、 pVg-RBS-ext 1
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3、 pVg-RBS-ext 2
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4、 sfp-RBS-ext 1
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5、 sfp-RBS-ext 2
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6、 PSB1C3回收
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9.14
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1、electrophoresis (ladder:1kb plus DNA)
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2、pVeg-RBS-ext2
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3、sfp-RBS-ext2
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4、pHT01 1
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5、pHT01 2
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9.15
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1、pfu: erP+BioBricks site PCR
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2、加A
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3、YerP 加A
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4、electrophoresis(1kb ladder,YerP1 bio,YerP2 bio)
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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)
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6、连接
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9.17
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1、transcribe: Bsub
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2、transcribe: T-YerP
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3、OEPCR1
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9.18
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1、枯草感受态检测——菌落数量
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2、回收pVeg+sfp OEPCR
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3、重叠PCR体系
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9.20
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1、OEPCR3
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2、菌落PCR(psb1C3-AH)
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3、菌PCR electrophoresis(ladder,YerP,sfp,GFP,negative control,positive control,6)
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4、回收1000kb左右的pvg-sfp
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5、回收OEPCR3
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9.21
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1、新backbone,BioBricks site用pfu和2017○34GP3
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9.22
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1、YerP PCR
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2、胶回收
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9.25
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1、PCR:backbone,BioBricks site
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9.26
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1、OEPCR3 加A
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2、electrophoresis(none,ladder D2000,BioBricks site,backbone)
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9.27
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1、electrophoresis: for testing the product of回收产物
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First: 2-5道:T-YerP○5 1-4; 6-9道:T-YerP○4 1-4)
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Second:(2-5道:AH,1-4;6-7道:BioBricks site,Backbone)
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2、transcribe pVeg-bfp
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9.28
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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)
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9.30
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1、菌液PCR
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2、Gibson片段-pveg-sfp PCR)
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3、electrophoresis(1,2,3,4,5,pveg sfp gibs,+,-)
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10.9
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1、回收Gib-(pveg-sfp)
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2、enzyme digestion pVeg-sfp
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3、ligate pSB1C3-pvg-sfp
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10.10
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1、electrophoresis(ladder,YerP1,YerP2,YerP3,YerP4,5‘ homologous arm,3‘ homologous arm,Gb,bb,BB)
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10.11
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1、electrophoresis(D2000,1,2,3,4,5,6,7,8,9,10)
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2、electrophoresis(D2000,11,12,13,14,15,16,17,18)
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10.16
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1、electrophoresis(ladder D2000,OEPCR3)
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2、transcribe(control,T-pveg sfp rbs,IDT’s ImrA)
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3、electrophoresis testing(1kb plus,OEPCR 回收产物,5‘,3‘,pSB1C3切, p SB1C3)
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10.18
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1、PCR菌(○1pSB1A3-GFPpSB1C3-pTet-GFP丙丁戊阴○2psb-pvg-sfp连接产物,OEPCR,RFP3,P3FG)
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10.19
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1、菌PCR(○1ladder 2000,sfp,GFP,negative control,positive control,6)(D2000,psb-pvg-sfp连接产物,OEPCR,RFP3,P3FG)
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10.21
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1、PCR
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2、enzyme digestion(psb1C3,RFP3,lmrA质粒,pvg-sfp)
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3、electrophoresis(ladder 1kb plus,OEPCR3,ImrA1,ImrA2,D1,E1)
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4、electrophoresis(ladder 1kb plus,A1,B1,C1)
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10.22
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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
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2、Ligate PSB1C3-lmrA
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3、 Transcribe PSB1C3-pveg-sfp转化得平板
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4、Gibson
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6、gov electrophoresis(1kb plus marker,lmrA切1,2回收,lmrA2*2,lmrA3*2,pvg sfp*2,PSB1C3线性,pSB1C3)
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3、菌PCR(D2000,浓pveg-sfp,OEPCR,阳)
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4、菌PCR(D2000,YerP,sfp)
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10.23
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1、electrophoresis(D2000, ImrA2,lmrA2,lmrA3,lmrA4)
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10.24
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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
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</p>
 
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Latest revision as of 03:59, 2 November 2017

RDFZ-China

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