Team:Paris/July 17

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Contents

1 Results of the transformations (Biobricks 2007)
2 Cloning of the transformations
3 Screening PCR of the transformations
- 3.1 Protocol
- 3.2 Results

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	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 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	12pb
55°C		n_oligoR	19 VR	v_oligoR	10µM 1µL	Sucess
Time Elongation		Water	23µL			Yes
1m30'		DNA	Toothpick of B0034			Image see bands 9 to 16
Number cycles						B0034
29						Ladder band 0

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