Team:Tokyo Metropolitan/Notebook/Pattern/2010/08/31

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Contents

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＞ Matsuura 2010-08-06 20hr 02min (10).JPG

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＞
Matsuura 2010-08-06 20hr 02min (11).JPG

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.

＜Result＞
File:Matsuura 2010-08-31 17hr 16min.tif

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＞
File:Matsuura 2010-08-31 21hr 56min.tif

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