Team:Evry/Notebook/w7

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Weeks

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16

Week 7: 23rd July - 29th July

Monday, 23rd June

Cloning:

4 different clones pCS2+ RFP => incubation in LB medium overnight at 37 degree celsius.

PCR:

PCR of pCS2+:
Reactants Volumes (µl)
GC Buffer 10
dNTPs 1
DNA 1
H2O 32,5
Primers (FW and RV) 2,5 each
fusion polymerase 0,5

Tuesday, 24th June

Plasmid Purification:

On the pCS2+ RFP clones incubated the day before:
DNA Concentration
Tube
Concentration (ng.uL-1)
1
144,2
2
161,9
3
184,9
4
120,7

Digestion:

Plasmid digestion
Tube
V DNA (uL)
V SpeI (uL)
V EcoRI (uL)
V buffer (uL)
V H2O (uL)
1
7
1
1
2
9
2
6,2
1
1
2
9,8
3
5,4
1
1
2
10,6
4
8,3
1
1
2
7,7

Gel migration:

Gel 0,8%

Thursday, 26th June

Friday, 27th June