Quality Control of Isothermal Amplified DNA Based on Short Tandem Repeat Analysis

Methods Mol Biol. 2015:1347:129-40. doi: 10.1007/978-1-4939-2990-0_10.

Abstract

This protocol describes the use of a 16plex PCR for the purpose assessing DNA quality after isothermal whole genome amplification (WGA). In short, DNA products, generated by amplification multiple displacement amplification, are forwarded to PCR targeting 15 short tandem repeats (STR) as well as amelogenin generating up to 32 different PCR products. After amplification, the PCR products are separated via capillary electrophoresis and analyzed based on the obtained DNA profiles. Isothermal WGA products of good DNA quality will result in DNA profiles with efficiencies of >90 % of the full DNA profile.

Keywords: Isothermal whole genome amplification; Quality control PCR; STR analysis; Single-cell analysis.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Computational Biology / methods
  • DNA*
  • Electrophoresis, Capillary
  • Genome
  • Genomics / methods
  • Genomics / standards
  • Microsatellite Repeats*
  • Multiplex Polymerase Chain Reaction / methods
  • Nucleic Acid Amplification Techniques / standards*
  • Quality Control*
  • Single-Cell Analysis / methods
  • Single-Cell Analysis / standards
  • Software

Substances

  • DNA