Team:Tokyo Metropolitan/Notebook/Pattern/2010/08/30
From 2010.igem.org
(→2:DNA Digestion) |
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<Sample, Materials><br /> | <Sample, Materials><br /> | ||
・PCR productions | ・PCR productions | ||
- | *[https://2010.igem.org/Team:Tokyo_Metropolitan/Project/Pattern/Protocol#Sample_Number/ | + | *[https://2010.igem.org/Team:Tokyo_Metropolitan/Project/Pattern/Protocol#Sample_Number/ 1L](8/25) 8μl |
- | *2L(8/26) 8μl | + | *[https://2010.igem.org/Team:Tokyo_Metropolitan/Project/Pattern/Protocol#Sample_Number/ 2L](8/26) 8μl |
- | *4H(8/26) 16μl | + | *[https://2010.igem.org/Team:Tokyo_Metropolitan/Project/Pattern/Protocol#Sample_Number/ 4H](8/26) 16μl |
- | *5H(8/26) 16μl | + | *[https://2010.igem.org/Team:Tokyo_Metropolitan/Project/Pattern/Protocol#Sample_Number/ 5H](8/26) 16μl |
・Digest enzyme | ・Digest enzyme |
Revision as of 09:48, 19 October 2010
Contents |
2010/08/30
1:PCR
<Member>
Mariko, Hitomi, Taka
<Sample>
・E-coli (having BBa_I13521 plasmid)
・E-coli (having BBa_K208017 plasmid)
・E-coli JM109 (NIPPON GENE)
<Protocol>
See Protocol 2
・Tube (temperature in annealing)
1.Promoter~signal (72.0℃)
3. mRFP~Terminator (69.0)
4. Promoter (70.0)
5. RBS~signal (70.0)
7. Terminator (69.0/67.5/66.0)
8. CRP (72.5)
Total 8 tubes.
2:DNA Digestion
<Member>
nito
<Sample, Materials>
・PCR productions
・Digest enzyme
- AvrⅡ
- NheⅠ
- SpeⅠ
<Protocol>
See Protocol 9
3:Electrophoresis
<Member>
Mariko, Taka
<Sample>
・PCR products
<Protocol>
SeeProtocol 8
4:Electrophoresis
<Member>
nito
<Sample>
- 1L(8/25)
- 2L(8/26)
- 4H(8/26)
- 5H(8/26)
(after digestion)
<Protocol>
SeeProtocol 8
And cut off gels included DNA.
5:DNA Ligation
<Member>
nito
<Sample>
- 1L(8/25)
- 2L(8/26)
- 4H(8/26)
- 5H(8/26)
(after digestion)
<Protocol>
See Protocol 3
We ligated 1 and 2, 4 and 5.
6:Electrophoresis
<Member>
nito
<Sample>
- 1L(8/25)
- 2L(8/26)
- 4H(8/26)
- 5H(8/26)
(after ligation)
<Protocol>
SeeProtocol 8