Verification of alleles by using peak height thresholds and quality control of STR profiling kits
2011 (English)In: Book of Abstracts, 2011, 134- p.Conference paper (Refereed)
In the autumn of 2010 SKL performed in-house validation of PowerPlex ESX 16 System (Promega). As the validationshowed that very low amounts of DNA (< 10 pg) may provide correct allele callings (peaks above 50 rfu),we investigated the linear range, i.e., the interval of DNA amounts where a profile is well balanced and doesnot contain drop-outs and/or drop-ins. The linear range as indicated by our results is approximately from 0.5ng (manufacturer’s recommendation) to 2.0 ng of DNA. Profiles generated by less than 0.5 ng contained intralocus imbalances and/or drop-outs. Above 2.0 ng “bleed through” occurs due to overload of template-DNA.A way to verify the correctness of a profile, without knowing anything about the condition of the template-DNA, is to use peak height thresholds adjusted to each marker and batch of kits used. SKL performs a qualitycontrol and adjust thresholds for each batch of kits. Three main tests are performed; detection limit, inhibitortolerance and signal repeatability. The detection limit is examined to identify at which concentration intralocus imbalances and drop-outs start to increase. The ability to overcome inhibition is checked by analysingvarying amounts of blood extracted with Chelex. Finally a set of replicates of control DNA is amplified (0.5 ngtemplate-DNA) to calculate the mean peak height and standard deviation at each locus. Generally, the peakheight thresholds vary between 200 and 250 rfu for heterozygote peaks. To verify allelic peaks below the setpeak height thresholds, SKL uses consensus analysis.
Place, publisher, year, edition, pages
2011. 134- p.
IdentifiersURN: urn:nbn:se:liu:diva-70271OAI: oai:DiVA.org:liu-70271DiVA: diva2:437704
24th World Congress of the International Society for Forensic Genetics 2011