Reduction of Powerplex(®) Y23 reaction volume for genotyping buccal cell samples on FTA(TM) cards.
PowerPlex(®) Y23 is a novel kit for Y-STR typing that includes new highly discriminating loci. The Israel DNA Database laboratory has recently adopted it for routine Y-STR analysis. This study examined PCR amplification from 1.2-mm FTA punch in reduced volumes of 5 and 10 μL. Direct amplification and washing of the FTA punches were examined in different PCR cycle numbers. One short robotically performed wash was found to improve the quality and the percent of profiles obtained. The optimal PCR cycle number was determined for 5 and 10 μL reaction volumes. The percent of obtained profiles, color balance, and reproducibility were examined. High-quality profiles were achieved in 90% and 88% of the samples amplified in 5 and 10 μL, respectively, in the first attempt. Volume reduction to 5 μL has a vast economic impact especially for DNA database laboratories.
DNA Database Laboratory, Division of Identification and Forensic Science (DIFS), Israel Police, National H.Q., Jerusalem, 91906, Israel.
Aliza
A
Raziel
Aviva
A
Dell'Ariccia-Carmon
Ashira
A
Zamir
D017046:Cost Savings; D016172:DNA Fingerprinting / Q000379:methods*; D005838:Genotype; D006801:Humans; D008297:Male; D018895:Microsatellite Repeats; D009061:Mouth Mucosa / Q000166:cytology*; D016133:Polymerase Chain Reaction / Q000379:methods*; D015203:Reproducibility of Results
DNA database; DNA typing; FTA cards; PowerPlex Y23; Y-short tandem repeats; forensic science; volume reduction