Difference between revisions of "Team:Paris Saclay/Notebook/July/10"
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| − | + | Preparation of Agarose Gel 1%, 0,5g in 50mL of 1X TAE, 0,5µL of BET | |
| + | Migration 0,06A 80V | ||
We confirm that the PCR was effective. We can continue the protocol. | We confirm that the PCR was effective. We can continue the protocol. | ||
Revision as of 10:46, 15 July 2015
Contents
Lab Work
PCR
by Coralie
PCR Mix (for 3 tubes)
- GC Buffer: 30µL
- dNTP 10mM: 3µL
- Forward Primer (dilution: 1/10e): 7,5µL
- Reverse Primer (dilution: 1/10): 7,5µL
- Template DNA K115017 (dilution 1/10e): 6µL
- DNA polymerase Phusion: 1,5µL
- H2O: 94,5µL
In each tube: 50µL from the mix
Cycle: Initiation: 98°C - 30seconds Cycle (30 repeats): 98°C - 10seconds / 53°C - 30seconds / 72°C - 10seconds Term.: 72°C - 5min Keep it at 4°C
Verification of PCR products by electrophoresis
by Coralie
Preparation of Agarose Gel 1%, 0,5g in 50mL of 1X TAE, 0,5µL of BET Migration 0,06A 80V
We confirm that the PCR was effective. We can continue the protocol.
Digestion
by Coralie
We digest 2 tubes of the PCR product Mix for each tube:
- XbaI: 1µL
- PstI: 1µL
- FastDigest Buffer: 2µL
- H2O: µL
- PCR product: 10µL
Incubation at 37°C for 1 hour
Purification of the digested PCR product
by Coralie
We use the Nucleospin kit from Magerey Nagel Keep the product at -20°C
Transformation :
by Johan, Seong Koo
- S03518
- B0030
- B0015
- K1399005
Members present:
- Instructors : Alice.
- Students : Johan, Pauline, Coralie, Audrey, Seong Koo.