	US 12,188,014 B1
	Compositions and methods for nucleic acid processing using blocking agents
Andrew D. Price, Hayward, CA (US); and Geoffrey McDermott, Livermore, CA (US)
Assigned to 10X GENOMICS, INC., Pleasanton, CA (US)
Filed by 10X GENOMICS, INC., Pleasanton, CA (US)
Filed on Jun. 6, 2019, as Appl. No. 16/434,102.
Claims priority of provisional application 62/703,303, filed on Jul. 25, 2018.
Int. Cl. C12N 15/10 (2006.01); C12N 9/22 (2006.01); C12N 15/11 (2006.01); C12Q 1/6806 (2018.01)

CPC C12N 15/1096 (2013.01) [C12N 9/22 (2013.01); C12N 15/11 (2013.01); C12Q 1/6806 (2013.01); C12N 2310/20 (2017.05); C12N 2800/80 (2013.01)]

27 Claims

1. A method for nucleic acid processing, comprising:

(a) providing a reaction mixture comprising:

(i) a transposase enzyme,

(ii) a first nucleic acid molecule comprising a target sequence,

(iii) a second nucleic acid molecule comprising a transposon end sequence, and

(iv) a third nucleic acid molecule comprising: (1) a first region comprising a blocking agent, wherein said blocking agent is not a uracil binding protein, and wherein said blocking agent prevents said transposase enzyme from interacting with said first region of said third nucleic acid molecule; and (2) a second region comprising a functional sequence capable of coupling to said first nucleic acid molecule;

(b) contacting said first nucleic acid molecule, said second nucleic acid molecule, and said transposase enzyme to generate a fourth nucleic acid molecule comprising said target sequence and said transposon end sequence of said second nucleic acid molecule; and

(c) contacting said third nucleic acid molecule and said fourth nucleic acid molecule to generate a fifth nucleic acid molecule comprising said target sequence, said transposon end sequence, and a sequence of said third nucleic acid molecule.