"
EPF-Lausanne/25 August 2009
From 2009.igem.org
Wet Lab
Miniprep:
Made according to the protocol of Aisgen Miniprep Kit Elution in 50 ul. The RD2 #5 plasmids prep as some that have already been prepared will be sent for sequencing this afternoon.
Mediums
New medium were made to grow the double-transformed DH5 a (LovTap BB and RO2) -> should be without thy.
M9 was redone, pH adjusted with amino acid and filtred. for 1L:
- 5g glucose
- 6g Na2PO4
- 3g KH2PO4
- 1g NH4Cl
- 0.5g NaCl
- 0.12g MgSO4
- 0.01g CaCl2
Trp Operon synthesis
Using the Klenow fragments protocol and primers:
- Trp operon Rev
- Trp operon Fwd
| 2x Trp | 1x Negative Control | |
|---|---|---|
|
NEB 2 (10x) |
2ul each |
2ul |
|
dNTP |
1ul each |
1ul each |
|
template primers |
1ul (RV) + 1.5ul (FW) each |
- |
|
Klenow enzyme |
1 ul each |
1 ul |
|
dH2O |
18.5 ul each |
21ul |
|
total |
25 ul each |
25 ul |
People in the lab
Nath, Basile, Gab, Christian
