From 2012.igem.org
Weeks
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