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1. (WO2017156218) METHOD OF MAKING POLYNUCLEOTIDES USING CLOSED-LOOP VERIFICATION

Pub. No.:    WO/2017/156218    International Application No.:    PCT/US2017/021498
Publication Date: Fri Sep 15 01:59:59 CEST 2017 International Filing Date: Fri Mar 10 00:59:59 CET 2017
IPC: C12Q 1/68
C12P 19/34
C40B 40/06
Applicants: PRESIDENT AND FELLOWS OF HARVARD COLLEGE
Inventors: CHURCH, George M.
GRISWOLD, Kettner John Frederick
Title: METHOD OF MAKING POLYNUCLEOTIDES USING CLOSED-LOOP VERIFICATION
Abstract:
A method for making a polynucleotide is provided including (a) delivering one or more reaction reagents including an error prone or template independent DNA polymerase, cations and a selected nucleotide to a reaction site including an initiator sequence having a terminal nucleotide for a time period and under conditions capable of covalently adding one or more of the selected nucleotide to the terminal nucleotide at the 3' end of the initiator such that the selected nucleotide becomes a terminal nucleotide, and (b) determining whether the selected nucleotide has been added to the terminal nucleotide, wherein if the selected nucleotide has not been added to the terminal nucleotide, then repeating step (a) until the selected nucleotide has been added, and (c) repeating steps (a) and (b) until the polynucleotide is formed.