Enhanced Adaptor Ligation
The present disclosure describes a method of adaptor ligation to the ends of the fragmented double-stranded DNA molecules.
1 . A composition comprising:
a polymerase possessing nick translation activity, a ligase and a partially double stranded adapter comprising oligonucleotide 1 and oligonucleotide 2; wherein oligonucleotide 1 comprises a 5′ phosphate; and wherein the 5′ portion of oligonucleotide 1 is complementary to the 3′ portion of oligonucleotide 2, and wherein the 3′ portion of oligonucleotide 1 is not complementary to the 5′ portion of oligonucleotide 2.
2 . The composition of claim 1 , wherein the polymerase is Taq polymerase or DNA Polymerase I and the ligase is E. coli DNA ligase or T4 DNA ligase.
3 . A composition comprising:
a polymerase possessing nick translation activity, a ligase and a partially double stranded adapter comprising oligonucleotide 1 and oligonucleotide 2; wherein oligonucleotide 1 lacks a 5′ phosphate; and wherein the 5′ portion of oligonucleotide 1 is complementary to the 3′ portion of oligonucleotide 2, and wherein the 3′ portion of oligonucleotide 1 is not complementary to the 5′ portion of oligonucleotide 2.