Paris/October 12

From 2007.igem.org

< Paris(Difference between revisions)
 
(3 intermediate revisions not shown)
Line 1: Line 1:
 +
[[Paris/October 11|yesterday]] -- [[Paris/October 13|tomorrow]] <br>
== PCR ==
== PCR ==
Line 4: Line 5:
<br>
<br>
------
------
-
Mi PCR :<br>
+
MIX PCR :<br>
- 27µL H2O<br>
- 27µL H2O<br>
- 10µL Buffer10X GCRich (Finnzyme)<br>
- 10µL Buffer10X GCRich (Finnzyme)<br>
Line 13: Line 14:
- 0,5µL Phusion Taq<br>
- 0,5µL Phusion Taq<br>
------
------
-
Timing : <br>
+
Timing : <br><br>
First :<br>
First :<br>
- 2' at 98°C<br>
- 2' at 98°C<br>
Line 22: Line 23:
- 10' at 72°C<br>
- 10' at 72°C<br>
- infinitas at 4°C
- infinitas at 4°C
 +
 +
== Digestion reactions ==

Latest revision as of 14:27, 15 October 2007

yesterday -- tomorrow

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.


MIX PCR :
- 27µL H2O
- 10µL Buffer10X GCRich (Finnzyme)
- 1,5µL DMSO 100%
- 2µL Template L59
- 4µL O47
- 4µL O48
- 0,5µL Phusion Taq


Timing :

First :
- 2' at 98°C

  • 30X :

- 20" at 98°C
- 1'40" at 72°C
then :
- 10' at 72°C
- infinitas at 4°C

Digestion reactions