Title:
TOUCHDOWN PCR DETECTION METHOD FOR DETECTING EXPRESSION LEVEL OF CIRCULAR RNA
Document Type and Number:
WIPO Patent Application WO/2024/032321
Kind Code:
A1
Abstract:
The present invention belongs to the field of molecular biology. A TouchDown PCR detection method for detecting the expression level of circular RNA. The method comprises: extracting total RNA from a human liver cancer cell line HepG2 cell, subjecting the total RNA to reverse transcription into cDNA, and carrying out real-time fluorescence quantitative detection on the circHDAC2 fragment by means of the TouchDown PCR program. Compared with common PCR amplification, the method has a better specificity and sensitivity to the amplification of circular RNA. The method reduces the difficulty of primer design, and can efficiently detect the expression level of circular RNA, laying a foundation for deep research into circular RNA in the future.
Inventors:
MENG CHUNYAN (CN)
LIU YUE (CN)
LONG YIFEI (CN)
LI XUEYING (CN)
LIU YANG (CN)
FENG FUMIN (CN)
LIU YUE (CN)
LONG YIFEI (CN)
LI XUEYING (CN)
LIU YANG (CN)
FENG FUMIN (CN)
Application Number:
PCT/CN2023/107804
Publication Date:
February 15, 2024
Filing Date:
July 18, 2023
Export Citation:
Assignee:
UNIV NORTH CHINA SCIENCE & TECHNOLOGY (CN)
International Classes:
C12Q1/6851; C12N15/11
Domestic Patent References:
| WO2022183011A1 | 2022-09-01 |
Foreign References:
| CN109609656A | 2019-04-12 | |||
| CN105018495A | 2015-11-04 |
Other References:
LIU, CHEN: "Optimization and Application of Rice Circular RNA Experiment Identification Method", CHINESE MASTER’S THESES FULL-TEXT DATABASE, AGRICULTURE SCIENCE AND TECHNOLOGY, 15 January 2019 (2019-01-15)
"Master's Thesis", 1 June 2020, NORTH CHINA UNIVERSITY OF SCIENCE AND TECHNOLOGY, CN, article XU, JIE: "Competitive Expression of Circ_0077692 and HDAC2 Affects Anti-tuberculosis Drug-induced Liver Injury through Histone Deacetylation", pages: 1 - 71, XP009552551, DOI: 10.27108/d.cnki.ghelu.2020.000034
ZHANG PEI, FAN YUAN, SUN XIAOPENG, CHEN LU, TERZAGHI WILLIAM, BUCHER ETIENNE, LI LIN, DAI MINGQIU: "profiling reveals universal molecular mechanisms responsive to drought stress in maize and Arabidopsis", THE PLANT JOURNAL, BLACKWELL SCIENTIFIC PUBLICATIONS, OXFORD., GB, vol. 98, no. 4, 1 May 2019 (2019-05-01), GB , pages 697 - 713, XP093137443, ISSN: 0960-7412, DOI: 10.1111/tpj.14267
DRULA RARES, BRAICU CORNELIA, CHIRA SERGIU, BERINDAN-NEAGOE IOANA: "Investigating Circular RNAs Using qRT-PCR; Roundup of Optimization and Processing Steps", INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES, MOLECULAR DIVERSITY PRESERVATION INTERNATIONAL (MDPI), BASEL, CH, vol. 24, no. 6, 16 March 2023 (2023-03-16), Basel, CH , pages 5721, XP093137452, ISSN: 1422-0067, DOI: 10.3390/ijms24065721
DATABASE Nucleotide 31 May 2023 (2023-05-31), ANONYMOUS : "Homo sapiens histone deacetylase 2 (HDAC2), transcript variant 2, non-coding RNA", XP093137457, retrieved from NCBI Database accession no. NR_033441.2
"Master's Thesis", 1 June 2020, NORTH CHINA UNIVERSITY OF SCIENCE AND TECHNOLOGY, CN, article XU, JIE: "Competitive Expression of Circ_0077692 and HDAC2 Affects Anti-tuberculosis Drug-induced Liver Injury through Histone Deacetylation", pages: 1 - 71, XP009552551, DOI: 10.27108/d.cnki.ghelu.2020.000034
ZHANG PEI, FAN YUAN, SUN XIAOPENG, CHEN LU, TERZAGHI WILLIAM, BUCHER ETIENNE, LI LIN, DAI MINGQIU: "profiling reveals universal molecular mechanisms responsive to drought stress in maize and Arabidopsis", THE PLANT JOURNAL, BLACKWELL SCIENTIFIC PUBLICATIONS, OXFORD., GB, vol. 98, no. 4, 1 May 2019 (2019-05-01), GB , pages 697 - 713, XP093137443, ISSN: 0960-7412, DOI: 10.1111/tpj.14267
DRULA RARES, BRAICU CORNELIA, CHIRA SERGIU, BERINDAN-NEAGOE IOANA: "Investigating Circular RNAs Using qRT-PCR; Roundup of Optimization and Processing Steps", INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES, MOLECULAR DIVERSITY PRESERVATION INTERNATIONAL (MDPI), BASEL, CH, vol. 24, no. 6, 16 March 2023 (2023-03-16), Basel, CH , pages 5721, XP093137452, ISSN: 1422-0067, DOI: 10.3390/ijms24065721
DATABASE Nucleotide 31 May 2023 (2023-05-31), ANONYMOUS : "Homo sapiens histone deacetylase 2 (HDAC2), transcript variant 2, non-coding RNA", XP093137457, retrieved from NCBI Database accession no. NR_033441.2
Attorney, Agent or Firm:
BEIJING FINELAND IP FIRM (CN)
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