"
Team:DTU-Denmark/Notebook/11 July 2013
From 2013.igem.org
(Difference between revisions)
(→Combine Nir operon with pZA21) |
(→Combine Nir operon with pZA21) |
||
| Line 9: | Line 9: | ||
===Combine Nir operon with pZA21=== | ===Combine Nir operon with pZA21=== | ||
| + | |||
| + | Following Mathildes Protocol for DpnI digestion of our linear pZA21 w. Uracil incerts | ||
Mastermix for 7 PCR tubes, Using the mastermix protocol we got from Mathilde. | Mastermix for 7 PCR tubes, Using the mastermix protocol we got from Mathilde. | ||
Revision as of 12:51, 11 July 2013
Contents |
208
Main purpose
- PCR with USER primers to combine Nir operon with pZA21
Who was in the lab
Ariadni,Henrike,Julia,Jakob
Procedure
Combine Nir operon with pZA21
Following Mathildes Protocol for DpnI digestion of our linear pZA21 w. Uracil incerts
Mastermix for 7 PCR tubes, Using the mastermix protocol we got from Mathilde.
triplets of each of the two Nir Operon extracts that we have made yesterday, including one negative control:
- Nir 2a(I,II,III)
- Nir 2b(I,II,III)
Using the following PCR program:
| time | Temp C | |
|---|---|---|
| Denaturing | 2 min | 98 |
| start of loop | 10 sec | 98 |
| annealing | 30 sec | 60 |
| elongation | 9 min | 72 |
| goto start of loop 35 times | ||
| hold | 10 |
