A cycle of chromosome replication must be initiated at a fixed time of C + D =60 minutes before cell division.

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#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.

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#Unit 13. Methods in Biology

Molecular beacons gives an idea for annealing.

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#Unit 13. Methods in Biology

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

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#Unit 13. Methods in Biology

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

of base substitution.

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#Unit 13. Methods in Biology

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

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#Unit 13. Methods in Biology

A modification of overlap extension PCR may also be used to construct directed mutations; this is termed megaprimer PCR .