Kulibaba R A, Liashenko Y V
Tsitol Genet. 2016 May-Jun;50(3):16-23.
The formation of different types of the artifacts in the amplification reaction with using different classes of the molecular-genetic markers (Indel and SSR) was studied. It was shown that DNA heteroduplexes formed during amplification of heterozygous samples, as fragments of target genes and microsatellite loci. During the electrophoresis in native polyacrylamide gel of the amplification products of homozygous samples for microsatellite loci it was revealed specific additional fragments that do not belong to the class of heteroduplex DNA. It was supposed, that the additional fragments belong to a special type of homoduplex DNA – non-linear homoduplexes. The analysis revealed that the formation of a non-linear homoduplex DNA takes place on the 20–25 cycle of the PCR, both at the amplification of the individual samples, and individual DNA fragments that was cut out from the gel.
研究了在使用不同类别的分子遗传标记(插入缺失和简单重复序列)进行扩增反应时不同类型假象的形成情况。结果表明,在杂合样本扩增过程中形成了DNA异源双链体,作为靶基因和微卫星位点的片段。在对微卫星位点纯合样本的扩增产物进行天然聚丙烯酰胺凝胶电泳时,发现了不属于异源双链DNA类别的特定额外片段。据推测,这些额外片段属于一种特殊类型的同源双链DNA——非线性同源双链体。分析表明,非线性同源双链DNA的形成发生在PCR的第20至25个循环,无论是在单个样本的扩增过程中,还是从凝胶中切出的单个DNA片段的扩增过程中。