Team:Tokyo Metropolitan/Notebook/Pattern/2010/08/31
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Contents |
2010/08/31
1:Electrophoresis
<Member>
Hitomi
<Sample>
PCR products.
<Protocol>
SeeProtocol 8
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
・Digest enzyme
- AvrⅡ
- NheⅠ
- SpeⅠ
<Protocol>
See Protocol 9
4:Electrophoresis
<Member>
nito, Hitomi
<Sample>
・PCR products
<Protocol>
SeeProtocol 8
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