Annealing Temperatures for PCR

Jayakumar, R R.Jayakumar at roswellpark.org
Thu Mar 30 09:34:45 EST 2006


 You can calculate it using several online tools. One of my favourite is
   http://www.basic.northwestern.edu/biotools/oligocalc.html
You can also search on google with keywords "annealing temperature
calculator tool".  The melting temperature is your annealing
temperature.

   But still the actual Annealing temperature has tto be optimized
through gradient PCR using different conc. Off Mg and salts.  The Tm
values only helps to give an idea.
Best of luck
Jayakumar

-----Original Message-----
From: methods-bounces at oat.bio.indiana.edu
[mailto:methods-bounces at oat.bio.indiana.edu] On Behalf Of
retnohh at mail.mitra.net.id
Sent: Wednesday, March 29, 2006 6:31 PM
To: methods at magpie.bio.indiana.edu
Subject: Annealing Temperatures for PCR

Dear sir,
 From internet I know that youa an expert in Molecular Biology research.
I have a problem with my student PCR.
If you do not mind, please help me to tell me how to calculate the exact
annealing primers for pair of primers:
5'- TTA GGG CAA GAG ATG GTA AGG -3'  and
5'- TTA TAA CAA TGA TGG AGG G -3'
Thank you very much for your kindness.

Best regard
Retnohh


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