Agilent Technologies, 11011 N. Torrey Pines Road, La Jolla, CA 92037, USA.
Methods. 2010 Apr;50(4):S15-8. doi: 10.1016/j.ymeth.2010.01.004.
The next-generation DNA sequencing workflows require an accurate quantification of the DNA molecules to be sequenced which assures optimal performance of the instrument. Here, we demonstrate the use of qPCR for quantification of DNA libraries used in next-generation sequencing. In addition, we find that qPCR quantification may allow improvements to current NGS workflows, including reducing the amount of library DNA required, increasing the accuracy in quantifying amplifiable DNA, and avoiding amplification bias by reducing or eliminating the need to amplify DNA before sequencing.
下一代 DNA 测序工作流程需要对要测序的 DNA 分子进行精确的定量，以确保仪器的最佳性能。在这里，我们展示了 qPCR 在下一代测序中用于定量 DNA 文库的应用。此外，我们发现 qPCR 定量可能会改进当前的 NGS 工作流程，包括减少所需文库 DNA 的量、提高可扩增 DNA 定量的准确性以及通过减少或消除测序前扩增 DNA 的需要来避免扩增偏倚。
