Team:Paris/August 4

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|colspan="8"|[[Image:KR000101_1.jpg|thumb|'''Gel : L100(1-8)''']]
|colspan="8"|[[Image:KR000101_1.jpg|thumb|'''Gel : L100(1-8)''']]
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==> '''Conclusion :''' we obtain a low quantity of PCR products, but at the good size.
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So now, we know that we can amplify one of the clones of L100 transformants bacteria.

Revision as of 15:21, 6 August 2008

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Contents

PCR Screening of Ligation Transformants

Use of 8 clones of Ligation transformants n°L100 for screening PCR


Protocol of screening PCR

  • Mix
Name Vol (µl) Concentration
Quick Load 25µl 2X
OligoF_VF2 (O18) 1µl 10µM
OligoR_VR (O19) 1µl 10µM
water 23µl


  • 50µl of Mix PCR by tube/clone
  • one toothpick of each clone's colony by tube
  • Program : Annealing 55°C - Time élongation 1'30" - Number cycle : 29


Conditions of electrophoresis

  • 10µl of ladder 1 kb
  • 20µl of screening PCR
  • migration ~30min at 100W on 0,8% gel


Results for L100

L100= D110 + D130 = RBS-tetR-ECFP-Ter Part icon rbs.pngIcon coding.pngPart icon rbs.pngPart icon reporter.pngPart icon terminator.png

Band 1-->8
Expected size 1042 pb
Measured size 1100 pb
Gel : L100(1-8)


==> Conclusion : we obtain a low quantity of PCR products, but at the good size. So now, we know that we can amplify one of the clones of L100 transformants bacteria.

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