Team:Paris/July 17

From 2008.igem.org

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* '''PCR : J23101 (A)'''
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* '''PCR 8 : J23101 (A)'''
{|border="1" Paris_PCR_8| Title = J23101 (A)
{|border="1" Paris_PCR_8| Title = J23101 (A)
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|v_oligoF
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|55°C
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[[Image:080717-5.png]]
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* '''PCR 9 : E0040 (A)'''
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[[Image:080717-6.png]]
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{|border="1" Paris_PCR_9| Title = E0040 (A)
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|'''PCRs setting'''
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|Quick Load
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|25µL
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|Expected size
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|Annealing
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|n_oligoF
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|18 VF2
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|v_oligoF
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|10µM 1µL
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|840pb
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|55°C
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|n_oligoR
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|19 VR
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|v_oligoR
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|10µM 1µL
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|Sucess
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|Time Elongation
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|Water
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|23µL
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|Yes
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|1m30'
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|DNA
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|Toothpick of E0040
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|Image see bands 4 to 5
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|-
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|Number cycles
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|[[Image:080717-7.png|thumb|E0040]]
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|29
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|Ladder band 0
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[[Image:080717-7.png]]
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* '''PCR 10 : R0040 (A)'''
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{|border="1" Paris_PCR_10| Title = R0040 (A)
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|'''PCRs setting'''
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|
 +
|Quick Load
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|25µL
 +
|
 +
|
 +
|Expected size
 +
|-
 +
|Annealing
 +
|
 +
|n_oligoF
 +
|18 VF2
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|v_oligoF
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|10µM 1µL
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|174pb
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|-
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|55°C
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|
 +
|n_oligoR
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|19 VR
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|v_oligoR
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|10µM 1µL
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|Sucess
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|-
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|Time Elongation
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|
 +
|Water
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|23µL
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|
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|
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|Yes
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|-
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|1m30'
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|
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|DNA
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|Toothpick of R0040
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|
 +
|
 +
|Image see bands 7 to 8
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|-
 +
|Number cycles
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|
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|
 +
|
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|
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|
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|[[Image:080717-7.png|thumb|R0040]]
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|-
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|29
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|Ladder band 0
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Revision as of 13:59, 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 0 : J23109 (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 J23109 Image see bands 1 to 8
Number cycles
J23109
29 Ladder band 0


  • PCR 1 : 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 2 : 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 3 : 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 4 : C0079 (K)
PCRs setting Quick Load 25µL Expected size
Annealing n_oligoF 18 VF2 v_oligoF 10µM 1µL 876pb
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


  • PCR 5 : R0079 (A)
PCRs setting Quick Load 25µL Expected size
Annealing n_oligoF 18 VF2 v_oligoF 10µM 1µL 277pb
55°C n_oligoR 19 VR v_oligoR 10µM 1µL Sucess
Time Elongation Water 23µL Yes
1m30' DNA Toothpick of R0079 Image see bands 9 to 16
Number cycles
R0079
29 Ladder band 0


  • PCR 6 : E1010 (K)
PCRs setting Quick Load 25µL Expected size
Annealing n_oligoF 18 VF2 v_oligoF 10µM 1µL 801pb
55°C n_oligoR 19 VR v_oligoR 10µM 1µL Sucess
Time Elongation Water 23µL Yes
1m30' DNA Toothpick of E1010 Image see bands 1 to 8
Number cycles
E1010
29 Ladder band 0


  • PCR 7 : B0034 (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 B0034 Image see bands 1 to 8
Number cycles
B0034
29 Ladder band 0


  • PCR 8 : J23101 (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 J23101 Image see bands 1 to 7
Number cycles
J23101
29 Ladder band 0


  • PCR 9 : E0040 (A)
PCRs setting Quick Load 25µL Expected size
Annealing n_oligoF 18 VF2 v_oligoF 10µM 1µL 840pb
55°C n_oligoR 19 VR v_oligoR 10µM 1µL Sucess
Time Elongation Water 23µL Yes
1m30' DNA Toothpick of E0040 Image see bands 4 to 5
Number cycles
E0040
29 Ladder band 0


  • PCR 10 : R0040 (A)
PCRs setting Quick Load 25µL Expected size
Annealing n_oligoF 18 VF2 v_oligoF 10µM 1µL 174pb
55°C n_oligoR 19 VR v_oligoR 10µM 1µL Sucess
Time Elongation Water 23µL Yes
1m30' DNA Toothpick of R0040 Image see bands 7 to 8
Number cycles
R0040
29 Ladder band 0
Retrieved from "http://2008.igem.org/Team:Paris/July_17"