TLS Online TPP Program

#Id: 2595

#Unit 13. Methods in Biology

Oligonucleotide-directed mutagenesis

The basic PCR mutagenesis system involves the use of two primary PCR reactions to produce two overlapping DNA fragments, both bearing the same mutation in the overlap region; this technique is thus termed overlap extension PCR .

TLS Online TPP Program

#Id: 2596

#Unit 13. Methods in Biology

Oligonucleotide-directed mutagenesis.

This technique requires a knowledge of nucleotide sequence, since an oligonucleotide may then be synthesized with the base mutation. 

Annealing of the oligonucleotide to complementary (except for the mutation) single-stranded DNA provides a primer for DNA polymerase to produce a new strand and thus incorporates the primer with the mutation.

TLS Online TPP Program

#Id: 2597

#Unit 13. Methods in Biology

Molecular beacons gives an idea for annealing.

TLS Online TPP Program

#Id: 2598

#Unit 13. Methods in Biology

TaqMan assay gives an idea for alleles in Real time PCR.

TLS Online TPP Program

#Id: 2599

#Unit 13. Methods in Biology

TaqMan assay also applicable in analyzing variations in the allele or deletion 

of base substitution.

TLS Online TPP Program

#Id: 2600

#Unit 13. Methods in Biology

Touch down PCR are derived to enhance primer binding in the annealing step.