PCR for NGS

Overview

Next-generation polymerases for next-generation sequencing

The Polymerase Chain Reaction (PCR) is acknowledged as one of the most enabling technologies in molecular biology.1 The advent of next-generation sequencing (NGS) paved the way for a plethora of new PCR applications. With the exception of PCR-free protocols (which have become the gold standard for human whole-genome sequencing), the majority of strategies used to convert samples to sequencing-ready NGS libraries involve one or more PCR steps. In addition, PCR-based methods are used at other stages of the sample prep process, including quality control and/or quantification steps. 

NGS brings many challenges to PCR technology. These include limited amounts of template, and low or variable sample quality. Library amplification steps require true (unbiased) replication of highly complex populations of molecules. Extremely high fidelity is essential to limit PCR errors that may result in false genetic variant calls. Whether you are generating amplicon libraries, amplifying or quantifying sequencing-ready libraries, or using PCR for a different purpose in your NGS sample preparation workflow, high amplification efficiency, low amplification bias and high fidelity are needed to process more samples successfully, get more information from every sample and optimize your sequencing resources. The directed evolution  platform employed at KAPA enabled us to develop next-generation DNA polymerases that are specifically engineered meet these challenges. 

Featured Applications

Roche Sample Prep Solutions offer a family of enzymes designed for robust amplification of different types of templates for a broad range of applications. KAPA enzymes have enabled life science and research for over a decade. Learn more about the benefits of our enzymes, and how researchers are using them for different purposes in NGS workflows.

  • NGS library amplification 
  • Somatic oncology research
  • Metagenomic and microbiome sequencing
  • Epigenomics
  • Single-cell RNA sequencing
  • CRISPR-Cas9 workflows
  • Amplicon sequencing
  • New and niche sequencing methods
  • NGS library quantification
  • Quality assessment of input DNA
  • Other PCR-based methods
  • High-fidelity amplification for cloning (incl. site-directed mutagenesis)

 

References:

  1. Bartlett JMS, Stirling D. et al. 2nd edition. Totowa: Humana Press; 2003. Preface. Available from: https://www.springer.com/us/book/9781592593842. Accessed November 2018.