How is heteroduplex DNA formed?

Heteroduplexes are formed during late stages of PCR reactions when high levels of products, such as in nested PCR (N-PCR) are generated, and other reactants such as primers and functional polymerase, are depleted. They are also formed by the deliberate mixing of separately amplified reactions.

What is Homoduplex DNA?

a DUPLEX DNA in which the two strands show COMPLEMENTARY BASE PAIRING throughout. Compare HETERODUPLEX.

How does Heteroduplex analysis work?

Hetero-duplex analysis is a low-medium high-throughput SNP detection technique. During heteroduplex analysis, the target gene is PCR amplified and the amplified products are then denatured, re-annealed slowly to form four different double-stranded DNA molecules from the two alleles of the gene (Paniego et al., 2015).

What is the Holliday model?

A conceptualization of general genetic recombination of newly replicated DNA during meiosis. An endonuclease nicks one strand of each of the two daughter duplexes at homologous sites which permits them to cross-over before being ligated.

Where are Holliday junctions found?

The Holliday junctions in homologous recombination are between identical or nearly identical sequences, leading to a symmetric arrangement of sequences around the central junction. This allows a branch migration process to occur where the strands move through the junction point.

How are heteroduplexes formed?

In brief, heteroduplexes are formed between different DNA alleles, for example, by mixing wild type and mutant amplified DNA fragments, followed by denaturation at 95°C and slow reannealing to room temperature ( White et al., 1992 ).

What is hetero-duplex analysis?

Hetero-duplex analysis is a low-medium high-throughput SNP detection technique. During heteroduplex analysis, the target gene is PCR amplified and the amplified products are then denatured, re-annealed slowly to form four different double-stranded DNA molecules from the two alleles of the gene (Paniego et al., 2015 ).

What is the difference between homoduplex and bulge Type heterod UPLEX?

If no mutation is present in the test DNA, then only a bulge type heteroduplex will be present, slightly retarded compared to the homoduplex.

What is the best diagnostic technique for short gels with heteroduplex?

A number of molecular diagnostic techniques, based on heteroduplex formation, are reported in the literature, such as enzymatic or chemical cleavage and DGGE, but HDA appears to be the most attractive one, as it can be performed rapidly on short gels without the need for specialized equipment and the use of radioactivity.

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