Patent ID: 9745614
Date: 2017-08-29
CPC Classifications: C12Q

Claim:
1. A method for generating a bisulfite converted library, the method comprising: a) ligating a first oligonucleotide sequence from a pool of oligonucleotide sequences to a 5′ end of a first polynucleotide from a plurality of polynucleotides from a sample to generate a first oligonucleotide-polynucleotide complex, b) ligating a second oligonucleotide sequence from the pool of oligonucleotide sequences to a 5′ end of a second polynucleotide from the plurality of polynucleotides to generate a second oligonucleotide-polynucleotide complex, wherein the second oligonucleotide sequence terminates at its 3′ end with one random base, wherein following the ligating of b), the one random base at the 3′ end of the second oligonucleotide sequence is immediately adjacent to the 5′ end of the second polynucleotide from the plurality of polynucleotides; c) ligating a third oligonucleotide sequence from the pool of oligonucleotide sequences to a 5′ end of a third polynucleotide from the plurality of polynucleotides to generate a third oligonucleotide-polynucleotide complex, wherein the third oligonucleotide sequence terminates at its 3′ end with two random bases, wherein following the ligating of c), a 3′ most base of the two random bases at the 3′ end of the third oligonucleotide sequence is immediately adjacent to the 5′ end of the third polynucleotide from the plurality of polynucleotides; d) ligating a fourth oligonucleotide sequence from the pool of oligonucleotide sequences to a 5′ end of a fourth polynucleotide from the plurality of polynucleotides to generate a fourth oligonucleotide-polynucleotide complex, wherein the fourth oligonucleotide sequence terminates at its 3′ end with three random bases, wherein following the ligating of d), a 3′ most base of the three random bases at the 3′ end of the fourth oligonucleotide sequence is immediately adjacent to a 5′ end of the fourth polynucleotide from the plurality of polynucleotides; e) treating the first, second, third, and fourth oligonucleotide-polynucleotide complexes with bisulfite, thereby generating a bisulfite converted polynucleotide library.