View source for VR/VF2 Gradient PCR Experiment

I tried gradient PCR to see if a higher annealing temperature would prevent the incorrect primer binding.

== Experiment ==
I prepared 8 reactions for both S03582 and I13033


'''PCR Reaction'''
*9 &mu;? PCR Supermix High Fidelity (Invitrogen)
*.25 &mu;L of 40 &mu;M VR
*.4 &mu;L of 25 &mu;M VF2
**''I ran out of 40 &mu;M VF2 so I used more of a less concentrated sample to make up the difference''
*1 &mu;L template DNA (~10 ng/&mu;L)


* Initial denature 95&deg;C - 5 min
* 35 cycles
** 94&deg;C - 30 sec
** Gradient from 54&deg;C to 70&deg;C - 30 sec
** 68&deg;C  - 36 sec
* Final extension 68&deg; - 10 min
* 4&deg;C forever


<nowiki>*</nowiki>The annealing temperature gradient ran from 54&deg;C to 70&deg;C, but due to the placement of 8-well strips samples were at the following annealing temperatures: 55.4&deg;C, 56.7&deg;C, 58.5&deg;C, 60.9&deg;C, 63.6&deg;C, 65.8&deg;C, 67.6&deg;C, 68.7&deg;C


I ran the results on a 1% Agarose gel for 30 minutes.

== Results ==
[[Image:070817.png|thumb|right|Results of Gradient PCR]]
The extra bands still appeared at the different annealing temperatures.