US Patent 12084715 Methods and systems for reducing artifactual antisense products

Disclosed herein is a method of preventing the amplification of artifactual antisense products when sequencing a target polynucleotide. Incorporation of modified nucleotides into handle polynucleotide sequences can allow for the selective degradation of artifact antisense products generated by mis-priming, prior to amplification, thus preventing them from appearing in the amplification library. Selective degradation can be accomplished using one or more agents configured to specifically cleave the handle polynucleotide at the modified nucleotide. Amplification can be performed using a primer that is configured to hybridize to the handle sequence and a second primer that is configured to hybridize to a complement of a target nucleotide.