Discussion
Which of the following is used to determine the appropriate stringency (Wallace rule)?
Answer: C
Correct answer: Tm = 4 x (No.of G:C base pair) + 2 x (No.of A:T base pair)
The Wallace rule for short oligonucleotides weights each base pair by the number of hydrogen bonds it makes - three for G:C, two for A:T - giving
\[ T_m = 4(G+C) + 2(A+T) \]in degrees Celsius. The heavier weight goes to G:C, which is the trap in the alternatives. The rule is a rough guide, reliable only for probes of roughly 14-20 bases.
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