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Ancient DNA

Volume 840 of the series Methods in Molecular Biology pp 121-132

Date:

Quantitative Real-Time PCR in aDNA Research

  • Michael BunceAffiliated withAncient DNA Laboratory, School of Biological Sciences and Biotechnology, Murdoch University Email author 
  • , Charlotte L. OskamAffiliated withAncient DNA Laboratory, School of Biological Sciences and Biotechnology, Murdoch University
  • , Morten E. AllentoftAffiliated withAncient DNA Laboratory, School of Biological Sciences and Biotechnology, Murdoch University

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Abstract

Quantitative real-time PCR (qPCR) is a technique that is widely used in the field of ancient DNA (aDNA). Quantitative PCR can be used to optimize aDNA extraction methodologies, to detect PCR inhibition, and to quantify aDNA libraries for use in high-throughput sequencing. In this chapter, we outline factors that need to be considered when developing efficient SYBR Green qPCR assays. We describe how to setup qPCR standards of known copy number and provide some useful tips regarding interpretation of qPCR data generated from aDNA templates.

Key words

qPCR Ancient DNA SYBR Green PCR inhibition qPCR standard DNA extraction optimization Library quantitation