Combinational usage of next generation sequencing and qPCR for the analysis of tumor samples

Methods. 2013 Jan;59(1):126-31. doi: 10.1016/j.ymeth.2012.11.002. Epub 2012 Nov 21.


The combination of multiple techniques especially those adding complementary information have proven to be beneficial in terms of data consistency. The employment of quantitative PCR (qPCR) prior to next generation sequencing (NGS) methods such as RNA-Seq and mutational analysis presented here does not only enhance data in terms of CNV integration and sample choice, but also allows a faster and more efficient workflow. Correct analysis of libraries prior to sequencing has proven to be a vital step for specific assumption and to some extent for a more parallel testing. By illustrating the combination of qPCR and NGS in oncological examples, the potential of this approach is presented.

Publication types

  • Review

MeSH terms

  • DNA Mutational Analysis / standards
  • Gene Library
  • High-Throughput Nucleotide Sequencing / standards*
  • Humans
  • Neoplasms / genetics*
  • Neoplasms / metabolism
  • Quality Control
  • RNA, Messenger / genetics
  • RNA, Messenger / metabolism
  • Real-Time Polymerase Chain Reaction / standards*
  • Sequence Analysis, RNA / standards


  • RNA, Messenger