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Team:Tokyo Metropolitan/Notebook/Pattern/2010/08/30
From 2010.igem.org
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
- 1L(8/25) 8μl
- 2L(8/26) 8μl
- 4H(8/26) 16μl
- 5H(8/26) 16μl
・Digest enzyme
- AvrⅡ
- NheⅠ
- SpeⅠ
＜Protocol＞
See Protocol 9
3：Electrophoresis
＜Member＞
Mariko, Taka
＜Sample＞
・PCR products
＜Protocol＞
SeeProtocol 8
＜Result＞
File:Matsuura 2010-08-30 15hr 17min.tif
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
＜Result＞
