US 12,252,732 B2
Methods and systems for nucleic acid preparation and chromatin analysis
Geoffrey McDermott, Livermore, CA (US)
Assigned to 10x Genomics, Inc., Pleasanton, CA (US)
Filed by 10x Genomics, Inc., Pleasanton, CA (US)
Filed on Jun. 21, 2023, as Appl. No. 18/339,082.
Application 18/339,082 is a continuation of application No. 16/854,475, filed on Apr. 21, 2020, granted, now 11,725,231.
Application 16/854,475 is a continuation of application No. PCT/US2018/057156, filed on Oct. 23, 2018.
Claims priority of provisional application 62/577,447, filed on Oct. 26, 2017.
Prior Publication US 2024/0052397 A1, Feb. 15, 2024
This patent is subject to a terminal disclaimer.
Int. Cl. C12Q 1/6806 (2018.01)
CPC C12Q 1/6806 (2013.01) 20 Claims
 
1. A method comprising:
(a) lysing a biological particle comprising chromatin comprising a native structure that includes a plurality of nucleosomes and one or more accessible regions positioned between the plurality of nucleosomes, wherein the lysing preserves the native structure of the chromatin;
(b) generating a plurality of template deoxyribonucleic acid (DNA) molecules from the one or more accessible regions of the native structure of the chromatin using a plurality of transposon-nucleic acid complexes;
(c) generating a plurality of ribonucleic acid (RNA) molecules from the plurality of template DNA molecules such that the plurality of RNA molecules corresponds to a plurality of positions of the plurality of nucleosomes of the chromatin; and
(d) generating a plurality of complementary DNA (cDNA) molecules from the plurality of RNA molecules using a plurality of template switching oligonucleotides such that the plurality of cDNA molecules corresponds to the plurality of positions of the plurality of nucleosomes of the chromatin.