METHODS, COMPOSITIONS, AND KITS FOR INHIBITING FORMATION OF ADAPTER DIMERS

Methods of reducing adapter-dimers in a sequencing library are provided according to aspects of the present disclosure, including: hybridizing excess 3′ adapters to blocker oligonucleotides forming a hybridized complex, such that the excess 3′ adapters are unavailable to form adapter-dimers, the hybridized complex including: a first blocker oligonucleotide 3′ terminus adjacent to an adenylated nucleotide at the 5′ terminus of a first unligated 3′ adapter and a second blocker oligonucleotide 3′ terminus adjacent to an adenylated nucleotide at the 5′ terminus of a second unligated 3′ adapter, wherein the first blocker oligonucleotide has a 5′ portion complementary to and hybridized to the first unligated 3′ adapter, the second blocker oligonucleotide has a 5′ portion complementary to and hybridized to the second unligated 3′ adapter, and the first blocker oligonucleotide has a 3′ portion complementary to, and hybridized to, a 3′ portion of the second blocker oligonucleotide.