How to calculate primer annealing temperature?

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veniesh
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How to calculate primer annealing temperature?

Post by veniesh » Tue Sep 29, 2009 3:34 pm

How to calculate primer annealing temperature? :idea: Please give the step and suitable formula (manual calculation)? :?: In my experiment DNA template will used and primer basepair between 18 to 25 basepair (Primer minimum size = 18 and Primer maximum size is 25). thanks for answer. :roll:

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JackBean
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Post by JackBean » Tue Sep 29, 2009 5:16 pm

I think, pDRAW32 program can calculate it ;)
On synthesis report I has found formula for primers of 13-50bp long:
Tm = 100.5 + (41*(yG+zC)/(wA+xT+yG+zC)) - (820/(wA+xT+yG+zC)) + 16.6*log10[Na+]
The last part adjusts the Tm for changes in the salt conc.

Try www.metabion.com/biocalc ;)

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