Team:Paris/July 17

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

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		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