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

Kód výsledku v IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F68081707%3A_____%2F03%3A17033153" target="_blank" >RIV/68081707:_____/03:17033153 - isvavai.cz</a>

Výsledek na webu

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DOI - Digital Object Identifier

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Jazyk výsledku

angličtina

Název v původním jazyce

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

Popis výsledku v původním jazyce

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 repetitivenature 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 homoduplexformation 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.

Název v anglickém jazyce

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

Popis výsledku anglicky

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 repetitivenature 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 homoduplexformation 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.

Druh

J_x - Nezařazeno - Článek v odborném periodiku (Jimp, Jsc a Jost)

CEP obor

BO - Biofyzika

OECD FORD obor

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Projekt

<a href="/cs/project/GA301%2F01%2F0590" target="_blank" >GA301/01/0590: Strukturní vlastnosti a prodlužování mononukleotidových a dinukleotidových mikrosatelitů lidského genomu</a><br>

Návaznosti

P - Projekt vyzkumu a vyvoje financovany z verejnych zdroju (s odkazem do CEP)<br>Z - Vyzkumny zamer (s odkazem do CEZ)

Rok uplatnění

2003

Kód důvěrnosti údajů

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů

Název periodika

Molecular Biology Reports

ISSN

0301-4851

e-ISSN

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Svazek periodika

30

Číslo periodika v rámci svazku

3

Stát vydavatele periodika

NL - Nizozemsko

Počet stran výsledku

9

Strana od-do

155-163

Kód UT WoS článku

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EID výsledku v databázi Scopus

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