Question

In a PCR reaction, following components were taken- ds DNA, Taq polymerase, dNTPs and ds primer, but even after 30 cycles, no amplification of the target DNA could be seen. The reason for this is because the  A. template is DNA and not RNA.  B. ddNTPs are not added in the reaction mixture.  C. primer is double stranded.  D. reverse transcriptase is not added.

Answer 1

The most likely reason for the lack of amplification in this PCR reaction is that the primer is double-stranded. PCR primers should be single-stranded in order to anneal to the target DNA and allow for the amplification to occur. Double-stranded primers are unable to anneal properly and cannot initiate the amplification process.