Team:Tokyo Metropolitan/Notebook/Pattern/2010/08/30

From 2010.igem.org

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(New page: {{:Team:Tokyo_Metropolitan/Header}} =2010/08/30= ==1:PCR == <Member><br /> Mariko, Hitomi, Taka <Sample><br /> ・E-coli (having BBa_I13521 plasmid) ・E-coli (having BBa_K20...)
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[[Image:Matsuura_2010-08-30_15hr_17min.tif]]
==4:Electrophoresis==
==4:Electrophoresis==

Revision as of 09:56, 22 September 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

  • 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>