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Rapid quantification of DNA libraries for next-generation sequencing

by: Bernd Buehler, Holly H. Hogrefe, Graham Scott, Harini Ravi, Carlos Pabón-Peña, Scott O’Brien, Rachel Formosa, Scott Happe
Methods, Vol. 50, No. 4. (April 2010), pp. S15-S18, doi:10.1016/j.ymeth.2010.01.004  Key: citeulike:6817276

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Abstract

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.


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