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Expansion during PCR of short single-stranded DNA fragments carrying nonselfcomplementary dinucleotide or trinucleotide repeats
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SYSNO ASEP 0127065 Document Type J - Journal Article R&D Document Type Journal Article Subsidiary J Ostatní články Title Expansion during PCR of short single-stranded DNA fragments carrying nonselfcomplementary dinucleotide or trinucleotide repeats Author(s) Reichová, Naďa (BFU-R)
Kypr, Jaroslav (BFU-R) RIDSource Title Molecular Biology Reports. - : Springer - ISSN 0301-4851
Roč. 30, č. 3 (2003), s. 155-163Number of pages 9 s. Language eng - English Country NL - Netherlands Keywords DNA ; PCR ; expansion Subject RIV BO - Biophysics R&D Projects GA301/01/0590 GA ČR - Czech Science Foundation (CSF) CEZ AV0Z5004920 - BFU-R Annotation We performed PCR of many DNA fragments of 6-32 nucleotides in length. Some of the fragments expanded into kilobase lengths even in the absence of the complementary strand. The dramatic expansion was observed for (CA)(8), (TG)(8), (CA)(4), (CA)(6), (CA)(12), (TG)(4), (TG)(6), (TG)(12), (TC)(10), (GA)(10) and other single strands. Similar expansions were exhibited by related trinucleotide repeats (TTG)(5), (CAA)(5), (TGG)(5), and (CCA)(5) as well. However even small perturbations of the strict repetitive nature of the DNA primary structure substantially reduced the expansions. The expansion products had properties characteristic for normal Watson-Crick duplexes. Hence either the Taq polymerase and/or other components of the PCR buffer promote homoduplex formation of the non-selfcomplementary fragments, which is necessary to prime the synthesis of the complementary DNA strand, or the Taq polymerase is able to copy the single-stranded DNA template without any priming effect. Workplace Institute of Biophysics Contact Jana Poláková, polakova@ibp.cz, Tel.: 541 517 244 Year of Publishing 2004
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