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Contents 1 2010/08/26 1.1 1：PCR 1.2 2：DNA extraction 1.3 3：DNA concentration measurement 1.4 4：Electrophoresis 1.5 5：Transformation with ECOS competent E-coli(NIPPON GENE)

2010/08/26

1：PCR

＜Member＞
Mariko, mizuki

＜Sample＞
・E-coli (having BBa_K208017 plasmid)

・E-coli (having BBa_I732901 plasmid)

・E-coli(JM109)

＜Protocol＞
See Protocol 2

・Tube(higher temperature/lower temperature in annealing)
2. JM109 (71.5℃/70.0)
4. Promoter(71.5/70.0)
5. RBS~signal (71.5/70.0)
6. lacZ (66.0/63.0)

Total 8 tubes.

2：DNA extraction

＜Member＞
nito

＜Sample＞
・PCR products

＜Protocol＞
SeeProtocol 4

3：DNA concentration measurement

＜Member＞
nito

＜Sample＞
・PCR products

＜Protocol＞

1. Add 2µl of TE and 2µl of sample into the tube, and mix it gently.
2. Fall in drops 1 on the measuring machine.

＜Result＞

DNA concentration

	concentration	A320	A260/A280	A260/A230
2H	95.5	14.84	2.011	0.054
2L	43.0	-0.025	1.792	0.203
4H	75.0	0.044	1.852	0.346
4L	63.0	-0.004	1.881	0.565
5H	35.0	2.96	2.000	0.160
5L	31.0	-0.020	1.873	0.071
6H	54.5	-0.078	1.946	0.215
6L	72.0	0.501	2.257	0.029

4：Electrophoresis

＜Member＞
nito

＜Sample＞
・PCR products

＜Protocol＞
SeeProtocol 8

＜Result＞
File:Matsuura 2010-08-26 16hr 14min.tif

5：Transformation with ECOS competent E-coli(NIPPON GENE)

＜Member＞
nito

＜Sample＞
・ECOS competent E-coli JM109(NIPPON GENE)

・Parts

• BBa_I13521
• BBa_B0010
• BBa_B0030
• BBa_J23110
• BBa_E0040

＜Protocol＞
SeeProtocol 7