From 2008.igem.org
(Difference between revisions)
|
|
| Line 249: |
Line 249: |
| | |Ladder band 0 | | |Ladder band 0 |
| | |} | | |} |
| | + | |
| | * '''PCR : pSB1A3 (A)''' | | * '''PCR : pSB1A3 (A)''' |
| | | | |
| Line 309: |
Line 310: |
| | |} | | |} |
| | | | |
| | + | |
| | + | * '''PCR : C0179 (A)''' |
| | + | |
| | + | {|border="1" Paris_PCR_0| Title = 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 |
| | + | | |
| | + | | |
| | + | | |
| | + | | |
| | + | | |
| | + | |[[Image:080717-2.png|thumb|C0179]] |
| | + | |- |
| | + | |29 |
| | + | | |
| | + | | |
| | + | | |
| | + | | |
| | + | | |
| | + | |Ladder band 0 |
| | + | |} |
| | | | |
| | | | |
Revision as of 13:06, 18 July 2008
|
← Yesterday ↓ Calendar ↑Tomorrow →
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
| 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
|
|
|
|
|
|
|
| 29
|
|
|
|
|
| Ladder band 0
|
| 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
|
|
|
|
|
|
|
| 29
|
|
|
|
|
| Ladder band 0
|
| 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
|
|
|
|
|
|
|
| 29
|
|
|
|
|
| Ladder band 0
|
| 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
|
|
|
|
|
|
|
| 29
|
|
|
|
|
| Ladder band 0
|
|
"
