Mar 25, 2014

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

BioRxiv : the Preprint Server for Biology
Castle RaleyDwight Nissley

Abstract

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.

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Mentioned in this Paper

Drug Carriers
NCOR2 wt Allele
Nucleic Acid Amplification Tests
Nucleic Acid Sequencing
Single Molecule Imaging
Gene Amplification Technique
Sequencing
Massively-Parallel Sequencing
Sequence Determinations, DNA
Plasma membrane-bound protein complex

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