Team:Paris/July 17

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Contents

Results of the transformations (Biobricks 2007)

Legende :

  • few , medium, a lot --> description of the number of CFU (Colony forming unit) produce by the transformation
  • A: Ampicilin, K: Kanamycin


Ref. Book Biobricks 2007 Antibio Results
- positive control A 2.800 CFU
- negative control A 0 CFU
2E_SP 1000 B0034 A few
10F_SP 1001 R0079 A medium
4F_SP 1013 C0079 K few
6H_SP 1001 C0179 A medium
6B_SP 1001 S03154 A 0 CFU
10F_SP 1002 J23101 A a lot
11F_SP 1002 J23109 A a lot
4C_SP 1000 R0040 A 2 CFU
2G_SP 1000 C0040 A 0 CFU
1C_SP 1008 PSB1A3 A few
7G_SP 1013 E1010 K medium
7E_SP 1001 E0040 A 2 CFU
2F_SP 1016 E0030 K a lot


Cloning of the transformations

  • Test 8 clones by Biobricks
  • Incubate O/N at 37°C on proper plates


Screening PCR of the transformations

Use of the same 8 clones of Biobricks that have been tested for cloning


Mix for PCR (x76 tubes)

25µl Quick load 2x 1µl Oligo F 1µl Oligo R 23µl H2O


Protocol

  • 50µl of Mix PCR by tubes
  • 1 clone
  • Programme : 105°C - for 29 cycles : 95°C; 5' - 95°C; 30" - 55°C; 30" - 72°C; 1,5'

Results

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PCRs setting Quick Load 25µL Expected size
Annealing n_oligoF 18 VF2 v_oligoF 10µM 1µL 35pb
55°C n_oligoR 19 VR v_oligoR 10µM 1µL Sucess
Time Elongation Water 23µL Yes
1m30' DNA Cycles= 29 n_oligoR= 19 VR v_oligoR= 10µM 1µL water= 23µL DNA= toothpick of J23109 Size expected= 35 pb Success= YES Image= Band=1-8
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