Evaluation of Direct PCR Amplification Using Various Swabs and Washing Reagents

J Forensic Sci. 2015 Nov;60(6):1542-52. doi: 10.1111/1556-4029.12865. Epub 2015 Sep 1.

Abstract

DNA profiles were generated via direct amplification from blood and saliva samples deposited on various types of swab substrates. Each of the six non-FTA substrates used in this research was punched with a Harris 1.2 mm puncher. After 0.1 μL of blood or 0.5 μL saliva, samples were deposited on each of these punches, samples were pretreated with one of four buffers and washing reagents. Amplification was performed using direct and nondirect autosomal and Y-STR kits. Autosomal and Y-STR profiles were successfully generated from most of these substrates when pretreated with buffer or washing reagents. Concordant profiles were obtained within and between the six substrates, the six amplification kits, and all four reagents. The direct amplification of substrates which do not contain lysing agent would be beneficial to the forensic community as the procedure can be used on evidence samples commonly found at crime scenes.

Keywords: DNA typing; direct amplification; forensic science; polymerase chain reaction; short tandem repeat; washing buffer.

MeSH terms

  • Blood Chemical Analysis
  • DNA Fingerprinting / methods*
  • Female
  • Humans
  • Indicators and Reagents*
  • Male
  • Microsatellite Repeats
  • Polymerase Chain Reaction*
  • Saliva / chemistry
  • Specimen Handling / instrumentation*

Substances

  • Indicators and Reagents