RT Journal Article
SR Electronic
T1 Preparation of next-generation DNA sequencing libraries from ultra-low amounts of input DNA: Application to single-molecule, real-time (SMRT) sequencing on the Pacific Biosciences RS II
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 003566
DO 10.1101/003566
A1 Castle Raley
A1 David Munroe
A1 Kristie Jones
A1 Yu-Chih Tsai
A1 Yan Guo
A1 Bao Tran
A1 Sujatha Gowda
A1 Jennifer L. Troyer
A1 Daniel R Soppet
A1 Claudia Stewart
A1 Robert Stephens
A1 Jack Chen
A1 TF Skelly
A1 Cheryl Heiner
A1 Jonas Korlach
A1 Dwight Nissley
YR 2014
UL http://biorxiv.org/content/early/2014/03/25/003566.abstract
AB We have developed and validated an amplification-free method for generating DNA sequencing libraries from very low amounts of input DNA (500 picograms – 20 nanograms) for singlemolecule sequencing on the Pacific Biosciences (PacBio) RS II sequencer. The common challenge of high input requirements for single-molecule sequencing is overcome by using a carrier DNA in conjunction with optimized sequencing preparation conditions and re-use of the MagBead-bound complex. Here we describe how this method can be used to produce sequencing yields comparable to those generated from standard input amounts, but by using 1000-fold less starting material.