E.coli Pattern Formation Project Notebook

August 2010 SUN MON TUE WED THU FRI SAT 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31

September 2010 SUN MON TUE WED THU FRI SAT 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30

October 2010 SUN MON TUE WED THU FRI SAT 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31

Contents 1 2010/08/31 1.1 1：Electrophoresis 1.2 2：PCR 1.3 3：DNA Digestion 1.4 4：Electrophoresis 1.5 5：Electrophoresis 1.6 6：DNA extraction 1.7 7：DNA extraction from gels 1.8 8：DNA Ligation 1.9 9：Electrophoresis

2010/08/31

1：Electrophoresis

＜Member＞
Hitomi

＜Sample＞
PCR products.

• 1 (8/25)
• 3 (8/26)
• 4 (8/26)
• 5 (8/26)
• 7 (8/30)
• 8 (8/30)

＜Protocol＞
SeeProtocol 8

＜Result＞

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)

1. Promoter~signal (72.0℃)
2. cyaA (71.5)
3. mRFP~Terminator (69.0)
4. Promoter (70.0)
5. RBS~signal (70.0)
6. lacZ (63.5)
7. Terminator (67.5)
8. 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＞

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＞