TLS Online TPP Program

#Id: 2544

#Unit 13. Methods in Biology

The efficiency of amplification declines with an increase in the length of the target sequence.

TLS Online TPP Program

#Id: 2545

#Unit 13. Methods in Biology

Efficiency of PCR is affected by primer length; it declines if the primers used for PCR arc too long.

TLS Online TPP Program

#Id: 2546

#Unit 13. Methods in Biology

The annealing temperature has a marked influence on PCR efficiency; a temperature higher than the ideal annealing temperature reduces PCR efficiency.

TLS Online TPP Program

#Id: 2547

#Unit 13. Methods in Biology

Primer sequence also· affects PCR efficiency. For example, if the primers used for PCR have complementary regions, primer dimers will be formed, and PCR efficiency will markedly decline.

TLS Online TPP Program

#Id: 2548

#Unit 13. Methods in Biology

The sequence of the target DNA itself can affect PCR efficiency. The GC-rich sequences in target DNA may form secondary structures in the single strands produced by denaturation; this could reduce PCR efficiency.

TLS Online TPP Program

#Id: 2549

#Unit 13. Methods in Biology

7-dcaza guanosine triphosphate (7-deaza OTP) can be used in place of OTP; it reduces hairpin loop formation without affecting the Watson-Crick base pairing required for annealing and primer extension.