US 12,467,074 B2
Method and apparatus for capturing high-purity nucleotides
Sung Hoon Kwon, Seoul (KR); Sung Sik Kim, Seoul (KR); Hui Ran Yeom, Cheorwon-gun (KR); Yong Hee Lee, Seoul (KR); Namphil Kim, Hanam-si (KR); Hamin Kim, Seoul (KR); and Wonjun Lee, Seoul (KR)
Assigned to SEOUL NATIONAL UNIVERSITY R&DB FOUNDATION, Seoul (KR)
Appl. No. 16/611,190
Filed by SEOUL NATIONAL UNIVERSITY R&DB FOUNDATION, Seoul (KR)
PCT Filed May 14, 2018, PCT No. PCT/KR2018/005493
§ 371(c)(1), (2) Date Nov. 5, 2019,
PCT Pub. No. WO2018/208133, PCT Pub. Date Nov. 15, 2018.
Claims priority of application No. 10-2017-0059558 (KR), filed on May 12, 2017.
Prior Publication US 2020/0165662 A1, May 28, 2020
Int. Cl. C12Q 1/6806 (2018.01); C12N 9/12 (2006.01); C12Q 1/686 (2018.01); C12Q 1/6869 (2018.01)
CPC C12Q 1/6806 (2013.01) [C12N 9/1252 (2013.01); C12Q 1/686 (2013.01); C12Q 1/6869 (2013.01); C12Q 2531/113 (2013.01); C12Q 2563/185 (2013.01)] 21 Claims
OG exemplary drawing
 
1. A method for capturing high-purity nucleic acid molecules, comprising (a) preparing nucleic acid fragments, (b) tagging the nucleic acid fragments with barcode sequences, (c) determining the location coordinates of sequence-validated ones of the nucleic acid fragments tagged with the barcode sequences, and (d) retrieving the sequence-validated fragments based on the location coordinates,
wherein the step (c) comprises:
step (c-1) of parallel sequencing all or some of the sequences of the fragments tagged with the barcode sequences in the step (b) to identify the barcode sequences of the sequence-validated fragments; and
step (c-2) comprising: i) immobilizing the fragments tagged with the barcode sequences in the step (b) on a support and ii) identifying the barcode sequences of the tagged fragments immobilized on the support by sequencing either simultaneously or sequentially to determine the location coordinates representing the barcode sequences identified by the sequence-validated fragments in the parallel sequencing step (c-1),
wherein, in step (c-2-i), the fragments tagged with the barcode sequences are directly attached to the support; the support is a patterned with repeated immobilization sites for immobilizing the fragments; each of the immobilization sites has a pillar structure and allows for the amplification of one fragment; and at least one of the immobilization sites includes only one fragment, and
wherein, in the step (c-2), the fragments tagged with the barcode sequences are amplified by rolling circle amplification (RCA).