Team:HokkaidoU Japan/Notebook/August24

From 2010.igem.org

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(Various Checks)
 
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Latest revision as of 07:44, 27 October 2010

Check to see if digestion visualization Primers Work

Electrophoresis after restriction enzyme digestion‎
  • All 4 of PCR products were purified via Microcon YM-10
Lane DNA
1 TSUDA Marker I
3 RBS
4 GFP
5 double terminator
6 Promoter

Promoter band in lane 6 wasn't visible

  • This vector didn't have a primer annealing site for our new primers

Check of Ligation Mixtures

TAKARABIO's Ligation Mixture [I] and Ligation Mixture Mighty Mix [M]

  • After PCR and gel extracted pSB1C3 volume of either Ligation Mixture equal to pSB1C3 solution volume was added
  • Transformed
  • incubated in 200 uL of LB medium
  • 100 uL of each solution was plated on mediums containing 34 ug/uL and 12 ug/uL of chloramphenicol

Various Checks

Electrophoresis to check if various DNA manipulations were successful
Lane DNA
2 λ/Hind III
3 Ligation between pSB1C3 vectors
4 Product obtained by using digestion visualization primer for 1-2N
5 Flow through of digestion visualization primer for 1-2N
6 λ/Hind III
7 Gel extracted pSB1C3

Band weren't visible in lane 3

  • Dimers, trimers and etc were hardly visible maybe severely diluted

In lane 7 had detectable by very weak band

  • There weren't any problems with gel extraction