When performing PCR, do you modify the Tm extension? I thought that the PCR extension Tm was always 72C...
Tm extension depends on the taq you are using for performing PCR.
that good quation
go to this wibsite
http://www.mutationdiscovery.com/md/MD.com/screens/optimase/OptimaseInput.html?action=none
and put your 2 primer
forward and reverse
and will show your tm
Tm depend on several factor
tag
number of nuclotide
length of primer
Yang is right. I use the two-step method with SYBR GREEN at 68 C.
C'est la températeure de polymérisation de la taq polymérase qui est toujours de 72°C.
Celle d'extention d'extension est varaible selon les primers: Elle est d'environ Tm-4.
Tu peux aussi utiliser un thermocycleur à gradient pour trouver expérimentalement ta températeure d'extension.
You can use any temperature where the polymerase works for extension. 72 is usually used for Taq as this is its optimal temperature, but many people use two step methods with a reannealing-extension step at temperatures as low as 60. You just have to add a few seconds to the extension time depending on the length of your target
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