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

From 2010.igem.org

Revision as of 06:05, 28 September 2010 by Mizuki (Talk | contribs)


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>
Matsuura 2010-08-06 20hr 02min (4).JPG