Team:Tokyo Metropolitan/Notebook/Pattern/2010/08/25

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Contents

2010/08/25

1：PCR

＜Member＞
Mariko, nito


＜Sample＞
・E-coli (having BBa_I13521 plasmid)

・E-coli (having BBa_K208017 plasmid)

・E-coli (having BBa_I732901 plasmid)


＜Protocol＞
See Protocol 1


・Tube(higher temperature/lower temperature in annealing)

  1. Promoter~signal (75.0℃/72.0)
  2. cyaA (76.5/75.0)
  3. mRFP~Terminator (72.5/71.0)
  4. Promoter (76.5/75.0)
  5. RBS~signal (76.5/75.0)
  6. lacZ (71.0/68.0)
  7. Terminator (71.0/68.0)
  8. CRP (72.5/71.0)

Total 16 tubes.

2： DNA extraction

＜Member＞
Mariko, nito, Hitomi


＜Sample＞
・PCR products


＜Protocol＞
See Protocol 4


3：DNA concentration measurement

＜Member＞
Mariko, nito, Hitomi


＜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
1H 165.0 0.142 1.886 0.821
1L 156.0 0.097 1.891 0.556
2H 13.5 0.059 1.868 0.341
2L 15.4 0.056 2.007 0.031
3H 0.7 0.55 -3.500 0.004
3L -4.8 1.02 1.484 -0.025
4H 10.0 0.055 1.905 0.035
4L 9.9 0.157 1.913 0.019
5H -41.0 9.39 1.464 3.475
5L 0.9 0.322 0.818 0.004
6H -36.0 5.55 1.636 0.321
6L 4.5 0.86 2.282 0.050
7H -29.5 6.07 1.439 2.070
7L -14.2 2.24 1.497 -0.062
8H 29.5 0.169 1.934 0.051
8L 26.5 0.081 1.978 0.038

4： Electrophoresis

＜Member＞
Mariko, nito, Hitomi


＜Sample＞
・PCR products


＜Protocol＞
See Protocol 8


＜Result＞
File:Matsuura 2010-08-25 19hr 43min.tif

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