Title:
TOPOLOGY CAPTURE-BASED RNA SAMPLE TREATMENT SYSTEM
Document Type and Number:
WIPO Patent Application WO/2023/241721
Kind Code:
A1
Abstract:
The present application discloses a cDNA synthesis method having improved high-throughput reverse transcription, template conversion and pre-amplification, so as to increase the yield and average length of cDNA libraries generated from single cells or single cell nuclei by means of high throughput. In the method and system, a large number of partitions (for example, a selective permeable compartment) are formed for performing single-cell or single-cell nucleus encapsulation, and the method and system allow single-operation or multi-operation chemical and/or biochemical treatment to be implemented within the partitions. Steps such as reverse transcription or full-length cDNA pre-amplification are performed within reaction compartment partitions, and fragmentation of a transposase complex-mediated full-length transcriptome amplification product is performed in each partition. This process retains library intermediate fragmented template DNA from same cells or cell nuclei in the selective permeable compartment. In addition, a partition (for example, a droplet) is secondarily formed by means of droplet microfluidics, and the selective permeable compartment and barcoded oligonucleotide microspheres of a fragmented full-length transcriptome amplification product from a single cell are encapsulated in the droplet, and the fragmented full-length transcriptome is labelled by means of the barcodes on the oligonucleotide microspheres.
Inventors:
ZHANG JINGWEI (CN)
LIU TIEMIN (CN)
LI TING (CN)
LIU TIEMIN (CN)
LI TING (CN)
Application Number:
PCT/CN2023/101050
Publication Date:
December 21, 2023
Filing Date:
June 19, 2023
Export Citation:
Assignee:
UNIV FUDAN (CN)
International Classes:
B01J13/20; C12P19/34; C12Q1/686
Foreign References:
| US20200400538A1 | 2020-12-24 | |||
| CN105579587A | 2016-05-11 | |||
| US20200131502A1 | 2020-04-30 | |||
| US20210214792A1 | 2021-07-15 |
Other References:
LI SIRAN, KENDALL JUDE, PARK SARAH, WANG ZIHUA, ALEXANDER JOAN, MOFFITT ANDREA, RANADE NISSIM, DANYKO CASSIDY, GEGENHUBER BRUNO, F: "Copolymerization of single-cell nucleic acids into balls of acrylamide gel", GENOME RESEARCH, COLD SPRING HARBOR LABORATORY PRESS, US, vol. 30, no. 1, 1 January 2020 (2020-01-01), US , pages 49 - 61, XP093117555, ISSN: 1088-9051, DOI: 10.1101/gr.253047.119
GRETA LEONAVICIENE ET AL.: "Multi-step processing of single cells using semi-permeable capsules", LAB CHIP., vol. 20, no. 21, 27 October 2020 (2020-10-27), XP055747240, DOI: 10.1039/D0LC00660B
GRETA LEONAVICIENE ET AL.: "Multi-step processing of single cells using semi-permeable capsules", LAB CHIP., vol. 20, no. 21, 27 October 2020 (2020-10-27), XP055747240, DOI: 10.1039/D0LC00660B
Attorney, Agent or Firm:
BEIJING TANGSONG IP FIRM (CN)
Download PDF:
Previous Patent: SYSTEM FOR PROCESSING SAMPLES AND METHOD FOR PREPARING SAME
Next Patent: METHODS AND SYSTEMS FOR IMAGE RECONSTRUCTION
Next Patent: METHODS AND SYSTEMS FOR IMAGE RECONSTRUCTION