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Expansion during PCR of short single-stranded DNA fragments carrying nonselfcomplementary dinucleotide or trinucleotide repeats

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    SYSNO ASEP0127065
    Document TypeJ - Journal Article
    R&D Document TypeJournal Article
    Subsidiary JOstatní články
    TitleExpansion during PCR of short single-stranded DNA fragments carrying nonselfcomplementary dinucleotide or trinucleotide repeats
    Author(s) Reichová, Naďa (BFU-R)
    Kypr, Jaroslav (BFU-R) RID
    Source TitleMolecular Biology Reports - ISSN 0301-4851
    Roč. 30, č. 3 (2003), s. 155-163
    Number of pages9 s.
    Languageeng - English
    CountryNL - Netherlands
    KeywordsDNA ; PCR ; expansion
    Subject RIVBO - Biophysics
    R&D ProjectsGA301/01/0590 GA ČR - Czech Science Foundation (CSF)
    CEZAV0Z5004920 - BFU-R
    AnnotationWe 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.
    WorkplaceInstitute of Biophysics
    ContactJana Poláková, polakova@ibp.cz, Tel.: 541 517 244
    Year of Publishing2004

Number of the records: 1