Question

A widely used method for calculating the annealing temperature for a primer used in PCR is 5 degrees below the melting temperature, Tₘ(°C), which is computed by the equation 81.5+0.41×(%GC)−(675/N), where %GC is the percentage of GC nucleotides in the oligonucleotide and N is the length of the oligonucleotide. Notice from the formula that both the GC content and the length of the oligonucleotide are variables. Assuming you have the following oligonucleotide as a primer,

5′-TTGAAAATATTTCCCATTGCC-3′

compute the annealing temperature for PCR. What is the relationship between and %GC? Why? (Note: In reality, this computation provides only a starting point for empirical determination of the most useful annealing temperature.) <>

Accepted Answer

Hello everyone here. We have a question asking us the temperature at which the primers bind to the DNA template is known as the blank. The temperature at which the primers bind to the DNA template is known as the kneeling temperature. And this is happening during the PcR cycle, also known as the polymerase chain reaction. The extension stage in which the preliminaries enzyme extends the primers by adding nucleotides to generate a new strand of DNA comes after the first maturation step where the double stranded DNA template is denatured into two distinct single stranded DNA molecules. So our answer here is C. A kneeling temperature, the temperature at which DNA strands unwind is termed the melting temperature. So B is incorrect and the temperature at which the enzyme activity is at its maximum is turning the optimum temperature, so A. Is incorrect. So again, our answer is C. A kneeling temperature. Thank you for watching. Bye.

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