Patent ID: 8932994
Filing Date: 2015-01-13
Classification: C12Q

Abstract:
1. A method of preparing a short insert library of adaptor-target-adaptor constructs, said method comprising: (a) preparing a plurality of short target polynucleotide duplex fragments having a distribution of sequences, wherein the short target polynucleotide duplex fragments average less than 150 base pairs in length; (b) treating the plurality of short target polynucleotide duplex fragments to phosphorylate the 5′ ends of each of the plurality of short target polynucleotide duplex fragments and to incorporate a single nucleotide overhang at each of the 3′ ends of each of the plurality of short target polynucleotide duplex fragments, wherein the treating produces a plurality of short, modified target polynucleotide duplex fragments wherein the plurality of short, modified target polynucleotide duplex fragments is purified, and wherein all steps in step (b) are performed at a temperature of less than 65° C.; and (c) ligating adaptor polynucleotides to both ends of each of the plurality of short, modified target polynucleotide duplex fragments to produce a short insert library of adaptor-target-adaptor constructs; wherein the distribution of sequences of the short target polynucleotide duplex fragments in the short insert library of adaptor-target-adaptor constructs is essentially equal to the distribution of sequences of the short target polynucleotide duplex fragments.