Rapid quantification of DNA libraries for next-generation sequencing.
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.
Buehler
Bernd
B
Agilent Technologies, 11011 N. Torrey Pines Road, La Jolla, CA 92037, USA.
Hogrefe
Holly H
HH
Scott
Graham
G
Ravi
Harini
H
Pabón-Peña
Carlos
C
O'Brien
Scott
S
Formosa
Rachel
R
Happe
Scott
S
D015723:Gene Library; D009693:Nucleic Acid Hybridization; D016133:Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D017422:Sequence Analysis, DNA / Q000379:methods*