Detection and quantitative characterization of artificial extra peaks following polymerase chain reaction amplification of 14 short tandem repeat systems used in forensic investigations

Electrophoresis
M Meldgaard, N Morling

Abstract

Detection on automated DNA sequencers of polymerase chain reaction (PCR) products of tetra- and penta-nucleotide short tandem repeat (STR) loci frequently reveals one or more extra peaks along with the true, major allele peak. The most frequent extra peak pattern is a single smaller peak which is one repeat unit shorter than the true allele peak. The existence of such artificial peaks is of special importance when the methods are used for forensic investigations because the artificial extra peaks may simulate true alleles when samples containing mixtures of DNA from different individuals are analyzed. We have investigated the relative levels of formation of extra peaks in 14 STR marker systems. We found that not only the parameters of the PCR but also factors determining the stringency during the post-PCR and pre-electrophoresis handling of samples were of importance for the formation of extra peaks. In our hands, the amounts of extra peaks were reduced (i) if the samples were effectively denatured immediately before loading, (ii) if they contained substantial amounts of formamide (i.e. > or = 50%), and (iii) if the temperature of the electrophoresis gel was above a certain level (i.e. > or = 43 degrees C). The results suggest ...Continue Reading

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Citations

Mar 11, 2011·Forensic Science International. Genetics·Clare BrookesJohn Buckleton
Aug 16, 2015·Forensic Science International. Genetics·Jessica Rothe, Marion Nagy
Sep 9, 2006·Electrophoresis·Marta Olejniczak, Wlodzimierz J Krzyzosiak
Mar 29, 2006·Journal of Forensic Sciences·Andreas P HellmannUwe Schleenbecker
May 14, 2011·Molecular Ecology Resources·E GuichouxR J Petit
Sep 15, 2019·International Journal of Legal Medicine·Yongsong ZhouBofeng Zhu
Jan 3, 2021·Forensic Science International. Genetics·Shota InokuchiNatsuko Mizuno
Dec 6, 2019·Forensic Science International. Genetics·Sarah RimanPeter M Vallone

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