Team:Paris/July 17

From 2008.igem.org

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===Results===
===Results===
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[[Image:Example.jpg]]
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{|Paris_PCR_0| Title = J23109 (A)
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|PCRs setting
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|
 +
|Quick Load
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|25µL
 +
|
 +
|Expected size
 +
|-
 +
|Annealing
 +
|
 +
|n_oligoF
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|18 VF2
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|v_oligoF
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|10µM 1µL
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|
 +
|35pb
 +
|-
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|55°C
 +
|
 +
|n_oligoR
 +
|19 VR
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|v_oligoR
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|10µM 1µL
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|
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|Sucess
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|-
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|Time Elongation
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|
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|Water
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|23µL
 +
|
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|Yes
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|-
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|1m30'
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|DNA
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|Cycles= 29
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|n_oligoR= 19 VR
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|v_oligoR= 10µM 1µL
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|water= 23µL
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|DNA= toothpick of J23109
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|Size expected= 35 pb
 +
|Success= YES
 +
|Image=
 +
|Band=1-8
 +
|}

Revision as of 11:25, 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


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

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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