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Team:Tokyo Metropolitan/Notebook/Pattern/2010/08/31
From 2010.igem.org
Contents |
2010/08/31
1：Electrophoresis
＜Member＞
Hitomi
＜Sample＞
PCR products.
＜Protocol＞
SeeProtocol 8
＜Result＞
.JPG)
2：PCR
＜Member＞
nito
＜Sample＞
・E-coli (having BBa_I13521 plasmid)
・E-coli (having BBa_K208017 plasmid)
・E-coli (having BBa_I732901 plasmid)
＜Protocol＞
See Protocol 2
・Tube (temperature in annealing)
- Promoter~signal (72.0℃)
- cyaA (71.5)
- mRFP~Terminator (69.0)
- Promoter (70.0)
- RBS~signal (70.0)
- lacZ (63.5)
- Terminator (67.5)
- CRP (72.5)
All tubes ×3. Total 24 tubes.
3：DNA Digestion
＜Member＞
Mariko, Hitomi
＜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
4：Electrophoresis
＜Member＞
nito, Hitomi
＜Sample＞
・PCR products
＜Protocol＞
SeeProtocol 8
＜Result＞
.JPG)
5：Electrophoresis
＜Member＞
Mariko, nito
＜Sample＞
- 1+2(8/31)
- 4+5(8/31)
(after digestion)
＜Protocol＞
SeeProtocol 8
And cut off gels included DNA.
6：DNA extraction
＜Member＞
nito
＜Sample＞
・PCR productions (8/31)
＜Protocol＞
SeeProtocol 4
7：DNA extraction from gels
＜Member＞
nito
＜Sample＞
- 1+2(8/31)
- 4+5(8/31)
(After electrophoresis)
＜Protocol＞
SeeProtocol 4
8：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.
9：Electrophoresis
＜Member＞
nito
＜Sample＞
- 1+2(8/31)
- 4+5(8/31)
(After ligation)
＜Protocol＞
SeeProtocol 8
＜Result＞
.JPG)
