Mutationist- Theories were advanced by some geneticists who observed that discretely different new phenotypes can arise by a process of mutation.

#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. 

#Id: 2550

#Unit 13. Methods in Biology

Addition of certain proteins, such as, bovine serum albumin enhances efficiency by protecting the DNA polymerase and by binding to PCR inhibitors. 

#Id: 2551

#Unit 13. Methods in Biology

5′ nuclease assay (TaqMan assay). PCR is undertaken with a fluorescence reporter/quencher pair (called RQ probe). 

#Id: 2552

#Unit 13. Methods in Biology

During extension by Taq polymerase, the probe is cleaved owing to the nuclease activity of the polymerase.

#Id: 2553

#Unit 13. Methods in Biology

Due to release of the reporter, there is a detectable increase in fluorescence, which is monitored in real-time PCR experiments.

#Id: 2587

#Unit 13. Methods in Biology

In case of nested PCR, the target sequence is amplified using a pair of PCR primers, and a portion of the amplification product is reamplified using another pair of PCR primers complementary to the regions located immediately beyond the 3'-ends of the first pair of primers.