US 12,391,975 B2
Systems and methods for transposon loading
Zahra Kamila Belhocine, Pleasanton, CA (US); and Jason Bell, Lafayette, LA (US)
Assigned to 10X GENOMICS, INC., Pleasanton, CA (US)
Filed by 10X Genomics, Inc., Pleasanton, CA (US)
Filed on Jul. 7, 2021, as Appl. No. 17/369,698.
Application 17/369,698 is a continuation of application No. PCT/US2020/017789, filed on Feb. 11, 2020.
Claims priority of provisional application 62/804,499, filed on Feb. 12, 2019.
Prior Publication US 2022/0170075 A1, Jun. 2, 2022
Int. Cl. C12Q 1/6806 (2018.01)
CPC C12Q 1/6806 (2013.01) 20 Claims
OG exemplary drawing
 
1. A method for processing a nucleic acid molecule, comprising:
(a) providing (I) a plurality of cells, nuclei, or cell beads, wherein a cell, nucleus, or cell bead of said plurality of cells, nuclei, or cell beads comprises said nucleic acid molecule and (II) a transposase molecule comprising a nucleic acid adapter, wherein said nucleic acid adapter comprises (i) a first single-stranded portion comprising a first nucleic acid sequence and (ii) a second single-stranded portion comprising a second nucleic acid sequence, wherein said first nucleic acid sequence is different from said second nucleic acid sequence, wherein said first single-stranded portion and said second single-stranded portion are in a nucleic acid loop;
(b) using said transposase molecule and said nucleic acid molecule to generate a nucleic acid fragment in said cell, nucleus, or cell bead, wherein said nucleic acid fragment (i) comprises a first end comprising said first nucleic acid sequence and said second nucleic acid sequence, (ii) comprises a second end comprising said first nucleic acid sequence and said second nucleic acid sequence, and (iii) is at least partially double stranded;
(c) after (b), partitioning, into a plurality of partitions, said plurality of cells, nuclei, or cell beads, wherein a partition of said plurality of partitions comprises (I) said cell, nucleus, or cell bead comprising said nucleic acid fragment and (II) a nucleic acid barcode molecule; and
(d) in said partition, cleaving said nucleic acid loop in said nucleic acid fragment to generate a processed nucleic acid fragment and reacting said nucleic acid barcode molecule with said processed nucleic acid fragment to generate a barcoded nucleic acid fragment.