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

From 2010.igem.org

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==6:DNA extraction==
==6:DNA extraction==

Revision as of 06:31, 28 September 2010


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>
Matsuura 2010-08-06 20hr 02min (14).JPG

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