Patent ID: 8741606
Filing Date: 2014-06-03
Classification: C12N,C12Q,G16B

Abstract:
1. A method of sequencing, comprising: a) appending a degenerate base region (DBR) to both ends of a plurality of target nucleic acid molecules in a genomic sample, wherein said DBR comprises a sequence comprising at least one nucleotide base selected from: R, Y, S, W, K, M, B, D, H, V, N and modified versions thereof and wherein said appending produces a population of asymmetrically tagged nucleic acid molecules, wherein each of the asymmetrically tagged nucleic acid molecules comprises: (i) a target polynucleotide, (ii) a first DBR sequence at the 5′ end of the target polynucleotide and (iii) a second DBR sequence at the 3′ end of the target polynucleotide, wherein: in each of said asymmetrically tagged nucleic acid molecules the first DBR sequence is different from the second DBR sequence; and the different asymmetrically tagged nucleic acid molecules in said population have different first DBR sequences and different second DBR sequences, relative to one another; b) amplifying at least some of said asymmetrically tagged nucleic acid molecules, thereby producing a population of amplified asymmetrically tagged nucleic acid molecules; and c) sequencing a plurality of the amplified asymmetrically tagged nucleic acid molecules to produce a plurality of sequences, wherein the sequencing step provides, for each of the asymmetrically tagged nucleic acid molecules that are sequenced: (i) the nucleotide sequence of at least a portion of a target polynucleotide; (ii) the nucleotide sequence of the first DBR sequence; and (iii) the nucleotide sequence of the second DBR sequence.