Paris/October 12
From 2007.igem.org
(Difference between revisions)
| Line 17: | Line 17: | ||
- 20" at 98°C<br> | - 20" at 98°C<br> | ||
- 1'40" at 72°C<br> | - 1'40" at 72°C<br> | ||
| + | then :<br> | ||
- 10' at 72°C<br> | - 10' at 72°C<br> | ||
- infinitas at 4°C | - infinitas at 4°C | ||
Revision as of 23:36, 14 October 2007
PCR
I made a PCR onto the L59 miniprep template having the [ptet>>lox66-GFP-terminator-lox71-mRFP] construction in order to add a forward restriction site for PacI and a backward restriction site for XmaI.
- 27µL H2O
- 10µL Buffer10X GCRich (Finnzyme)
- 1,5µL DMSO 100%
- 2µL Template L59
- 4µL O47
- 4µL O48
- 0,5µL Phusion Taq
- 2' at 98°C
- 30X :
- 20" at 98°C
- 1'40" at 72°C
then :
- 10' at 72°C
- infinitas at 4°C
