Overcoming a barrier for DNA polymerization in triplex-forming sequences.

V. N. Potaman, John Bissler

Research output: Contribution to journalArticle

13 Citations (Scopus)

Abstract

Folded structures in the DNA template, such as hairpins and multi-stranded structures, often serve as pause and arrest sites for DNA polymerases. DNA polymerization is particularly difficult on mirror-repeated homopurine.homopyrimidine templates where triple-stranded (triplex) structures may form between the nascent and folded template strands. In order to use a linear PCR amplification approach for the structural analysis of DNA in mirror-repeated sequences we modified a conventional protocol. The barrier for DNA synthesis can be eliminated using an oligonucleotide that hybridizes with the template to prevent its folding and is subsequently displaced by the progressing polymerase. The described approach is potentially useful for sequencing and analysis of chemical adducts and point mutations in a variety of sequences prone to the formation of folded structures, such as long hairpins and quadruplexes.

Original languageEnglish (US)
JournalNucleic Acids Research
Volume27
Issue number15
StatePublished - Jan 1 1999
Externally publishedYes

Fingerprint

Polymerization
DNA
DNA-Directed DNA Polymerase
Point Mutation
Oligonucleotides
Polymerase Chain Reaction

All Science Journal Classification (ASJC) codes

  • Genetics

Cite this

Overcoming a barrier for DNA polymerization in triplex-forming sequences. / Potaman, V. N.; Bissler, John.

In: Nucleic Acids Research, Vol. 27, No. 15, 01.01.1999.

Research output: Contribution to journalArticle

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