公开(公告)号：US20230141527A1

公开(公告)日：2023-05-11

申请号：US17809540

申请日：2022-06-28

Applicant: GUARDANT HEALTH, INC.

Inventor： Andrew KENNEDY ,  Stefanie Ann Ward MORTIMER ,  AmirAli TALASAZ ,  Darya CHUDOVA ,  Helmy ELTOUKHY ,  Oliver ZILL ,  Richard B. LANMAN ,  Rebecca NAGY ,  Christine LEE ,  Kimberley BANKS

IPC: C12Q1/6806 ,  C12Q1/6855 ,  C12Q1/6886

CPC classification number: C12Q1/6806 ,  C12Q1/6855 ,  C12Q1/6886 ,  C12Q2600/156 ,  C12Q2600/158

Abstract: Methods of preparing double-stranded nucleic acids with single-stranded overhangs for amplification and sequencing are disclosed. Contacting a blunt-ended double-stranded nucleic acid molecules with Taq results in non-templated directed addition of a single nucleotide to the 3′ ends of the nucleic acid with A added most frequently followed by G followed by C and T. G tailing is sufficiently frequent that the efficiency of ligation of nucleic acid molecules to adapters can be significantly increased by including adapters tailed with T and C. The ligation efficiency can be increased even further with blunted-ended adapters to ligate to blunt-ended nucleic acid molecules that failed to undergo tailing.