Team:UAB-Barcelona/PCRN

From 2009.igem.org

(Difference between revisions)
(PCR of nitrite detection sequences)
Line 12: Line 12:
Marker, 3x49ºC, controls, marker, 3x52ºC, controls
Marker, 3x49ºC, controls, marker, 3x52ºC, controls
 +
 +
From left side to right: lambda-DNA hinIII; 3 x nirk 49ºC; control without primers; control without DNA template; lambda-DNA hinIII; 3 x nirk 52ºC; control without primers; control without DNA template; lambda-DNA hinIII
'''Annealing temperature'''
'''Annealing temperature'''
Line 19: Line 21:
https://static.igem.org/mediawiki/2009/f/f3/Pnsr.jpg
https://static.igem.org/mediawiki/2009/f/f3/Pnsr.jpg
   
   
-
Marker, 4x65ºC, controls, marker, 4x68ºC, controls
+
lambda-DNA hinIII; 4 x nsr 65ºC; control without DNA template; control without primers; lambda-DNA hinIII; 4 x nsr 68ºC; control without DNA template; control without primers; lambda-DNA hinIII

Revision as of 02:15, 22 October 2009

Encap raro.jpg Documento sin título

PCR of nitrite detection sequences

We used GoTaq Green Master Mix (Promega)12

Annealing temperature

nirk (270 pb): as lowest temperature is 60ºC, the temperature we should set in the thermociclator would be 55ºC. But it didn't work properly at this temperature, so we set the temperature to 52ºC and 49ºC. PCR in both cases worked properly

NirK.jpg

Marker, 3x49ºC, controls, marker, 3x52ºC, controls

From left side to right: lambda-DNA hinIII; 3 x nirk 49ºC; control without primers; control without DNA template; lambda-DNA hinIII; 3 x nirk 52ºC; control without primers; control without DNA template; lambda-DNA hinIII

Annealing temperature

nsr(500 pb): the temperature is 76ºC, so we made the PCR at 71ºC. It didn't work. But when we tried with 65 and 68ºC it worked properly.

Pnsr.jpg

lambda-DNA hinIII; 4 x nsr 65ºC; control without DNA template; control without primers; lambda-DNA hinIII; 4 x nsr 68ºC; control without DNA template; control without primers; lambda-DNA hinIII