Question

In humans, congenital heart disease is a common birth defect that affects approximately 1 out of 125 live births. Using reverse transcription PCR (RT-PCR) Samir Zaidi and colleagues [(2013) Nature 498:220.223] determined that approximately 10 percent of the cases resulted from point mutations, often involving histone function. To capture products of gene expression in developing hearts, they used oligo(dT) in their reverse transcription protocol.

How would such a high %T in a primer influence annealing temperature?

Accepted Answer

Hello everyone here. We have a question that says the optimal and kneeling temperature for a pcr reaction is determined by a measuring the template DNA is melting temperature. This is incorrect. We need the temperature of the primers, not the template D. N. A. Be evaluating a variety of temperatures to identify the one that yields the most specific and efficient amplification. This is correct. But let's look at our other two, increasing the kneeling temperature until no amplification products are created. This will result in non specific amplification or no amplification. So C is incorrect and D will do the same, decreasing the kneeling temperature until no amplification products are created. So D is also incorrect. So our answer here is be evaluating a variety of temperatures to identify the one that yields the most specific and efficient amplification. Thank you for watching. Bye.

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