Team:Paris/July 17

From 2008.igem.org

(Difference between revisions)
(→Results)
(→Results)
Line 129: Line 129:
===Results===
===Results===
-
* '''PCR : J23109'''
+
* '''PCR : J23109 (A)'''
{|border="1" Paris_PCR_0| Title = J23109 (A)
{|border="1" Paris_PCR_0| Title = J23109 (A)
Line 189: Line 189:
|}
|}
 +
 +
* '''PCR : E0030 (K)'''
 +
 +
{|border="1" Paris_PCR_0| Title = E0030 (K)
 +
|'''PCRs setting'''
 +
|723pb
 +
|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 E0030
 +
|
 +
|
 +
|Image see bands 9 to 16
 +
|-
 +
|Number cycles
 +
|
 +
|
 +
|
 +
|
 +
|
 +
|[[Image:080717-2.png|thumb|J23109]]
 +
|-
 +
|29
 +
|
 +
|
 +
|
 +
|
 +
|
 +
|Ladder band 0
 +
|}
[[Image:080717-2.png]]
[[Image:080717-2.png]]

Revision as of 12:43, 18 July 2008

← Yesterday

↓ Calendar ↑

Tomorrow →



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

  • 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 723pb 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 E0030 Image see bands 9 to 16
Number cycles
J23109
29 Ladder band 0

080717-2.png

080717-3.png

080717-4.png

080717-5.png

080717-6.png

080717-7.png

Retrieved from "http://2008.igem.org/Team:Paris/July_17"