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

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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＞
Matsuura 2010-08-06 20hr 02min (7).JPG

4：Electrophoresis

＜Member＞
nito


＜Sample＞

(after digestion)


＜Protocol＞
SeeProtocol 8

And cut off gels included DNA.

＜Result＞
Matsuura 2010-08-06 20hr 02min (8).JPG

5：DNA Ligation

＜Member＞
nito


＜Sample＞

(after digestion)


＜Protocol＞
See Protocol 3

We ligated 1 and 2, 4 and 5.

6：Electrophoresis

＜Member＞
nito


＜Sample＞

(after ligation)


＜Protocol＞
SeeProtocol 8


＜Result＞
Matsuura 2010-08-06 20hr 02min (9).JPG

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