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1. (WO2018161019) METHODS FOR OPTIMIZING DIRECT TARGETED SEQUENCING

Pub. No.:    WO/2018/161019    International Application No.:    PCT/US2018/020744
Publication Date: Sat Sep 08 01:59:59 CEST 2018 International Filing Date: Sat Mar 03 00:59:59 CET 2018
IPC: C12P 19/34
C12Q 1/68
C12Q 1/6869
C40B 20/00
C40B 30/04
Applicants: COUNSYL, INC.
Inventors: LAI, Henry
CHU, Clement
Title: METHODS FOR OPTIMIZING DIRECT TARGETED SEQUENCING
Abstract:
Described are methods for selecting an amount of a critical parameter (such as an amount of a sequencing library, amount of a capture probe library, or a number of amplification cycles) for direct targeted sequencing. The methods include hybridizing capture probes in a capture probe library to surface-bound oligonucleotides; extending the surface-bound oligonucleotides using the hybridized capture probes as a template; hybridizing nucleic acid molecules from a sequencing library to the surface-bound capture probes; extending the surface-bound capture probes using the hybridized nucleic acid molecules as a template; amplifying the surface-bound complements of the nucleic acid molecules by bridge amplification for a number of amplification cycles; sequencing the amplified surface-bound complements of the nucleic acid molecules to determine an average cluster density after a predetermined number of sequencing cycles; repeating these steps at a plurality of different amounts of the critical parameter; and selecting an amount of the critical parameter.