Team:Paris/August 4

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PCR Screening of the 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= D106 + D120 = RBS-tetR-ECFP-Ter Part icon rbs.pngIcon coding.pngPart icon reporter.pngPart icon terminator.png

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