Title:
MICRO CELL CHIP METHOD
Document Type and Number:
WIPO Patent Application WO/2019/153852
Kind Code:
A1
Abstract:
A micro cell ChIP method, the method comprising: 1). dividing cell samples to be detected into a number n of groups, n being a non-zero natural number; 2). crosslinking and fixing the nth group sample; 3). processing the nth group sample using cell membrane punch agent; 4). using Tn5 transposases for enzyme digestion to interrupt the chromatin fragments of the nth group sample and connecting barcode sequences and primer sequences at two ends of the product fragment DNA, when n≥2, the barcode sequences and/or primer sequences used in each group of step 4) being different; 5). when n≥2, combining the various groups of samples; and 6). gathering DNA fragments combined with target protein and using the primer as index for database building, and carrying out sequencing analysis. The method has high database building efficiency during sequencing, can realize site capturing to transcription factor of infinitesimal cells, and can ensure better efficiency, stability and accuracy.
Inventors:
HE AIBIN (CN)
AI SHANSHAN (CN)
LI CHEN (CN)
AI SHANSHAN (CN)
LI CHEN (CN)
Application Number:
PCT/CN2018/118433
Publication Date:
August 15, 2019
Filing Date:
November 30, 2018
Export Citation:
Assignee:
UNIV BEIJING (CN)
International Classes:
C12Q1/68; C12Q1/6804
Domestic Patent References:
| WO2016169431A1 | 2016-10-27 | |||
| WO2017084023A1 | 2017-05-26 | |||
| WO2017034970A1 | 2017-03-02 |
Foreign References:
| CN108315387A | 2018-07-24 | |||
| CN104818331A | 2015-08-05 | |||
| CN103966317A | 2014-08-06 | |||
| CN103853936A | 2014-06-11 | |||
| CN106834208A | 2017-06-13 |
Other References:
SCHMIDL C. ET AL.: "ChIPmentation: Fast, Robust, Low-Input ChIP-seq for Histones and Transcription Factors", NATURE METHODS, vol. 12, no. 10, 17 August 2015 (2015-08-17) - October 2015 (2015-10-01), pages 963 - 965, XP055566448, doi:10.1038/nmeth.3542
ROTEM A. ET AL.: "Single- cell ChIP-seq Reveals Cell Subpopulations Defined by Chromatin State", NATURE BIOTECHNOLOGY, vol. 33, no. 11, 12 October 2015 (2015-10-12), pages 1165 - 1172, XP055235766, doi:10.1038/nbt.3383
ROTEM A. ET AL.: "High-Throughput Single- Cell Labeling (Hi-SCL) for RNA-Seq Using Drop-Based Microfluidics", PLOS ONE, vol. 10, no. 5, 22 May 2015 (2015-05-22), pages e0116328, XP055227089
ROTEM A. ET AL.: "Single- cell ChIP-seq Reveals Cell Subpopulations Defined by Chromatin State", NATURE BIOTECHNOLOGY, vol. 33, no. 11, 12 October 2015 (2015-10-12), pages 1165 - 1172, XP055235766, doi:10.1038/nbt.3383
ROTEM A. ET AL.: "High-Throughput Single- Cell Labeling (Hi-SCL) for RNA-Seq Using Drop-Based Microfluidics", PLOS ONE, vol. 10, no. 5, 22 May 2015 (2015-05-22), pages e0116328, XP055227089
Attorney, Agent or Firm:
CHOFN INTELLECTUAL PROPERTY (CN)
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