Robust Sub-nanomolar Library Preparation for High Throughput Next Generation Sequencing.

ABSTRACT:

Background

Current library preparation protocols for Illumina HiSeq and MiSeq DNA sequencers require ≥2 nM initial library for subsequent loading of denatured cDNA onto flow cells. Such amounts are not always attainable from samples having a relatively low DNA or RNA input; or those for which a limited number of PCR amplification cycles is preferred (less PCR bias and/or more even coverage). A well-tested sub-nanomolar library preparation protocol for Illumina sequencers has however not been reported. The aim of this study is to provide a much needed working protocol for sub-nanomolar libraries to achieve outcomes as informative as those obtained with the higher library input (≥ 2 nM) recommended by Illumina's protocols.

Results

Extensive studies were conducted to validate a

SUBMITTER: Wu WW 

PROVIDER: S-EPMC5935984 | biostudies-literature | 2018 May

REPOSITORIES: biostudies-literature