Team:Paris/July 17

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(Difference between revisions)
(→Results)
(→Screening PCR of the transformations)
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Use of the same 8 clones of Biobricks that have been tested for cloning  
Use of the same 8 clones of Biobricks that have been tested for cloning  
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===Mix for PCR (x76 tubes)===
 
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25µl Quick load 2x
 
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1µl Oligo F
 
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1µl Oligo R
 
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23µl H2O
 

Revision as of 13:13, 18 July 2008

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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


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

  • PCR : J23109 (A)
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 Toothpick of J23109 Image see bands 1 to 8
Number cycles
J23109
29 Ladder band 0


  • PCR : E0030 (K)
PCRs setting Quick Load 25µL Expected size
Annealing n_oligoF 18 VF2 v_oligoF 10µM 1µL 723pb
55°C n_oligoR 19 VR v_oligoR 10µM 1µL Sucess
Time Elongation Water 23µL Yes
1m30' DNA Toothpick of E0030 Image see bands 9 to 16
Number cycles
E0030
29 Ladder band 0


  • PCR : pSB1A3 (A)
PCRs setting Quick Load 25µL Expected size
Annealing n_oligoF 18 VF2 v_oligoF 10µM 1µL
55°C n_oligoR 19 VR v_oligoR 10µM 1µL Sucess
Time Elongation Water 23µL Yes
1m30' DNA Toothpick of pSB1A3 Image see bands 1 to 8
Number cycles
pSB1A3
29 Ladder band 0


  • PCR : C0179 (A)
PCRs setting Quick Load 25µL Expected size
Annealing n_oligoF 18 VF2 v_oligoF 10µM 1µL
55°C n_oligoR 19 VR v_oligoR 10µM 1µL Sucess
Time Elongation Water 23µL Yes
1m30' DNA Toothpick of C0179 Image see bands 9 to 16
Number cycles
C0179
29 Ladder band 0


  • PCR : C0079 (K)
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 Toothpick of C0079 Image see bands 1 to 8
Number cycles
C0079
29 Ladder band 0


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