What is the relation between Ta and Tm of the primers in PCR reactions? ?

2009-01-27 3:15 pm
I have read some readings online and is Ta should be 5 degree below Tm?
Does Tm stand for melting temperature of the DNA?
what does this formula: Tm = 4(G + C) + 2(A + T)oC ?? is the G, C , A, T the amount of the bases in the primers?
Thank you , urgently need help

回答 (1)

2009-01-27 3:33 pm
✔ 最佳答案
The Tm is the melting temperature. The Ta is the optimum annealing temperature (for use in your annealing step of the PCR reaction). Yes it is usually around 5 degrees below the Tm (although this varies considerably and I would always advise a temperature gradient PCR to determine the actual optimum annealing temperature for your primers).
The A, T, C and G in that formula refers to the number of time the bases are present in your primer sequence and it will give you an approximate theroretical Tm. If you then subtract 5 you will get an approximate theoretical Ta.
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