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

PCR Screening of the transformants

Use of the same 8 clones of Biobricks that have been tested for cloning

Protocol

• 50µl of Mix PCR by tubes
• 1 clone
• Program : 105°C - for 29 cycles : 95°C; 5' - 95°C; 30" - 55°C; 30" - 72°C; 1,5'

Results

• PCR 1 : J23109 (A)

PCRs setting		Quick Load	25µL			Expected size
Annealing		n_oligoF	18 VF2	v_oligoF	10µM 1µL	1000pb
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 2 : 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 3 : 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 4 : 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 5 : 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 6 : 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 7 : 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 8 : 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 7
Number cycles						B0034
29						Ladder band 0

• PCR 9 : J23101 (A)

PCRs setting		Quick Load	25µL			Expected size
Annealing		n_oligoF	18 VF2	v_oligoF	10µM 1µL	1000pb
55°C		n_oligoR	19 VR	v_oligoR	10µM 1µL	Sucess
Time Elongation		Water	23µL			Yes
1m30'		DNA	Toothpick of J23101			Image see bands 2 to 7
Number cycles						J23101
29						Ladder band 0

• PCR 10 : 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 11 : 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