Methods for increasing yield of sequencing libraries

A method for producing a sequencing library comprising:

• providing a plurality of double stranded symmetric modified target nucleic acids comprising a first adapter sequence at each end, wherein the first adapter sequence comprises a DNA lesion, with the proviso that the first adapter does not comprise a universal molecular identifier;
• extending the modified target nucleic acids with a lesion-intolerant polymerase to generate a plurality of single stranded asymmetric modified target nucleic acids comprising at the 5′ end of each strand the first adapter sequence and at the 3′ end of each strand the complement of a portion of the first adapter;
• annealing a primer to the plurality of single stranded asymmetric modified target nucleic acids, the primer comprising from 5′ to 3′ a second adapter sequence and an annealing domain, the annealing domain comprising a nucleotide sequence that anneals to the complement of the portion of the first adapter of the plurality of single stranded asymmetric modified target nucleic acids;
• extending the 3′ end of the annealed asymmetric modified target nucleic acids with a lesion-intolerant polymerase using the primer as template,
• wherein the extending results in a plurality of asymmetric modified target nucleic acids comprising from 5′ to 3′ (i) the first adapter, (ii) the target nucleic acid, (iii) the complement of the portion of the first adapter, and (iv) the complement of the second adapter.