DNA typing of a polymerase chain reaction amplified D1S80/amelogenin multiplex using capillary electrophoresis and a mixed entangled polymer matrix

Alice R. Isenberg, Bruce R. McCord, Barbara W. Koons, Bruce Budowle, Ralph O. Allen

Research output: Contribution to journalArticle

40 Scopus citations

Abstract

In this study, a technique was developed to separate by capillary electrophoresis (CE) the widely varying DNA fragment sizes produced by a multiplex polymerase chain reaction (PCR) amplification of the loci D1S80 and amelogenin. Experiments were performed to analyze different buffer systems and obtain optimal resolution for the separation. A matrix composed of two different molecular weights of the same polymer was constructed to separate the DNA fragments with baseline resolution, and a cubic spline fit was used to estimate the size of DNA fragments over 350 base pairs. Over 100 samples were examined to demonstrate the rapid, robust and precise characteristics of this CE system. An average relative standard deviation of 0.3% was obtained for the sizing of the D1S80 alleles in these samples. DNA from mixed body fluid samples, samples subjected to environmental insult, and D1S80 sequence variants were also typed successfully. These results demonstrate that CE is a viable method for analysis of D1S80 and amelogenin forensic DNA samples.

Original languageEnglish
Pages (from-to)1505-1511
Number of pages7
JournalELECTROPHORESIS
Volume17
Issue number9
DOIs
StatePublished - 1 Sep 1996

Keywords

  • Amelogenin
  • Capillary electrophoresis
  • D1S80
  • Forensic
  • Laser-induced fluorescence
  • Polymerase chain reaction

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