PT - JOURNAL ARTICLE AU - Castle Raley AU - David Munroe AU - Kristie Jones AU - Yu-Chih Tsai AU - Yan Guo AU - Bao Tran AU - Sujatha Gowda AU - Jennifer L. Troyer AU - Daniel R Soppet AU - Claudia Stewart AU - Robert Stephens AU - Jack Chen AU - TF Skelly AU - Cheryl Heiner AU - Jonas Korlach AU - Dwight Nissley TI - 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 AID - 10.1101/003566 DP - 2014 Jan 01 TA - bioRxiv PG - 003566 4099 - http://biorxiv.org/content/early/2014/03/25/003566.short 4100 - http://biorxiv.org/content/early/2014/03/25/003566.full 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.