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＞

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.

＜Result＞

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＞