Compositions and methods for generating massively parallel nucleic acid sequencing libraries

Abstract

Oligonucleotide primers and methods of use in producing sequencing libraries are provided according to aspects of the present disclosure which include, from 5' to 3′, a homopolymer-hybridizing region, and an anchor region comprising 5′-(λ)N-3′, wherein the homopolymer-hybridizing region is a contiguous sequence of 5 to 20 elements, wherein the elements are nucleotides or Tincreasing nucleotide analogs, wherein at least 4 of the elements are Tincreasing nucleotide analogs, wherein the homopolymer-hybridizing region hybridizes to a complementary homopolymer tract of a target nucleic acid, wherein the complementary homopolymer tract comprises a contiguous sequence of complementary elements, wherein X is any nucleotide or nucleotide analog with the proviso that λ is not a nucleotide or nucleotide analog complementary to a complementary element of the complementary homopolymer tract, and wherein N is any nucleotide or nucleotide analog.