US 12,344,876 B2
Spatially addressable control of polymerase activity
Bichlien Hoang Nguyen, Seattle, WA (US); and Jake Smith, Seattle, WA (US)
Assigned to Microsoft Technology Licensing, LLC, Redmond, WA (US)
Filed by MICROSOFT TECHNOLOGY LICENSING, LLC, Redmond, WA (US)
Filed on Oct. 30, 2020, as Appl. No. 17/086,055.
Prior Publication US 2022/0136021 A1, May 5, 2022
Int. Cl. C12P 19/34 (2006.01); C12N 9/12 (2006.01)
CPC C12P 19/34 (2013.01) [C12N 9/1252 (2013.01); C12Y 207/07007 (2013.01)] 20 Claims
 
1. A method for enzymatic synthesis of polynucleotides comprising:
(a) incubating an array having a plurality of initiators attached thereto with a reaction reagent solution comprising template-independent polymerase and with a selected species of nucleotide, wherein the reaction reagent solution has an unsuitable pH for the template-independent polymerase; and
(b) altering pH conditions at a selected location on the array such that activity of the template-independent polymerase is changed, without changing availability of a metal cofactor complexed with the template-independent polymerase,
wherein altering the pH conditions comprises altering pH conditions to an optimum pH for the template-independent polymerase thereby increasing activity of the template-independent polymerase at the selected location, and
wherein altering the pH conditions at the selected location on the array further comprises (i) delivering an acid or a base to the selected location with a targeted fluid deposition instrument, or (ii) exposing a photoacid or photobase in the reaction reagent solution to a light source.