Question

Most of the techniques described in this chapter (blotting, cloning, PCR, etc.) are dependent on hybridization (annealing) between different populations of nucleic acids. Length of the strands, temperature, and percentage of GC nucleotides weigh considerably on hybridization. Two other components commonly used in hybridization protocols are monovalent ions and formamide. A formula that takes monovalent Na⁺ ions ((M[Na⁺]) and formamide concentrations into consideration to compute a Tₘ (temperature of melting) is as follows:

Tₘ=81.5+16.6(log M[Na+])+0.41(%GC)−0.72(%formamide)

For the following concentrations of Na⁺ and formamide, calculate the Tₘ. Assume 45% GC content.
[Na⁺] % Formamide
0.825 20
0.825 40
0.165 20
0.165 40

Accepted Answer

Hello everyone here. We have a question asking us to identify the correct sequence of different steps of the polymerase chain reaction. Our first step is denature duration and during DNA duration the two strands of the double stranded DNA template are separated by heating it to a high temperature. Our 2nd step is a kneeling and this is when the temperature is dropped to a predetermined and healing temperature, which enables the primers to o'neill to their complementary sequences on the single stranded DNA template. And lastly we have extension. And this is when the primers are extended by adding nucleotides to form a new strain of D. N. A. And this is done at the temperature. That is best for the D. N. A polyamorous enzyme being employed. So our answer here is C. DNA melting primer binding primer extension. Thank you for watching. Bye.

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