"
Team:Heidelberg/Notebook/Sensing Group/Notebook/10thweek
From 2008.igem.org
(Difference between revisions)
(→Monday, 10/06/2008) |
(→Tuesday, 10/07/2008) |
||
| Line 129: | Line 129: | ||
== Tuesday, 10/07/2008 == | == Tuesday, 10/07/2008 == | ||
| + | * Repetition of Mutagenesis PCR with Phusion and same programme, but 30 cycles | ||
| + | --> only LuxS2 positive --> Gelextraction | ||
| + | |||
| + | |||
| + | *New PCR with Turbo Pfu and iProof (18 cycles) | ||
| + | ** 5(10) µl buffer | ||
| + | ** 1 µl forward Primer (10 µM stock) | ||
| + | ** 1 µl reverse Primer | ||
| + | ** 1 µl dNTPs | ||
| + | ** 2 µl Template | ||
| + | ** 39(34) µl water | ||
| + | ** 1 µl Polymerase | ||
| + | ** 30 sec @ 95 °C ||30 sec @ 95 °C | 60 sec @ 55 °C | 30 sec @ 68 °C || 8 min @ 68 °C | 4°C hold | ||
== Wednesday, 10/08/2008 == | == Wednesday, 10/08/2008 == | ||
Revision as of 19:32, 26 October 2008
Contents |
Monday, 10/06/2008
- Mutagenesis PCR of LuxS,P with Phusion and the following programme (20 cycles)
- PCR programme: 30 sec @ 98°C || 10 sec @ 98 °C | 30 sec @ 55 °C | 30 sec @ 72 °C || 5 min @ 72 °C | 4°C
--> No PCR product
Tuesday, 10/07/2008
- Repetition of Mutagenesis PCR with Phusion and same programme, but 30 cycles
--> only LuxS2 positive --> Gelextraction
- New PCR with Turbo Pfu and iProof (18 cycles)
- 5(10) µl buffer
- 1 µl forward Primer (10 µM stock)
- 1 µl reverse Primer
- 1 µl dNTPs
- 2 µl Template
- 39(34) µl water
- 1 µl Polymerase
- 30 sec @ 95 °C ||30 sec @ 95 °C | 60 sec @ 55 °C | 30 sec @ 68 °C || 8 min @ 68 °C | 4°C hold
