Title:
MITOCHONDRIAL GENOMIC LIBRARY BASED ON HIGH-THROUGHPUT SEQUENCING AND METHOD FOR CONSTRUCTING THE LIBRARY
Document Type and Number:
WIPO Patent Application WO/2017/193832
Kind Code:
A1
Abstract:
A mitochondrial genomic library based on high-throughput sequencing and a method for constructing the library. The method comprises the following steps: S1. performing a one-step PCR amplification of total mitochondrial DNA from total DNA; S2. fragmenting the total mitochondrial DNA to obtain a DNA fragment; S3. performing end repair on the DNA fragment to obtain a blunt-end DNA fragment; S4. A-tailing a 3' end of the blunt-end DNA fragment to obtain an A-tailed DNA fragment; S5. adding a linker on the 3' end of the A-tailed DNA fragment to obtain a DNA fragment with linkers; S6. performing LM-PCR on the DNA fragment with linkers; and S7. performing PCR on an LM-PCR product. The method for constructing the mitochondrial genomic library performs PCR amplification on total DNA to obtain total mitochondrial DNA, thereby ensuring accurate amplification of a human mitochondrial genome and improving the accuracy and precision of mutation detection by preventing contamination or mixing of a homologous sequence from a human genome.
Inventors:
ZHANG WEI VICTOR (CN)
Application Number:
PCT/CN2017/082495
Publication Date:
November 16, 2017
Filing Date:
April 28, 2017
Export Citation:
Assignee:
GUANGZHOU JIAJIAN MEDICAL TESTING CO LTD (CN)
International Classes:
C12N15/10; C12Q1/68; C40B40/06; C40B50/06
Foreign References:
| CN105907748A | 2016-08-31 | |||
| CN103173441A | 2013-06-26 | |||
| CN103571854A | 2014-02-12 | |||
| CN104099349A | 2014-10-15 | |||
| CN103602735A | 2014-02-26 | |||
| CN103614482A | 2014-03-05 | |||
| CN105112499A | 2015-12-02 | |||
| CN103103624A | 2013-05-15 | |||
| CN105087767A | 2015-11-25 |
Other References:
LIU, LI ET AL.: "The application of ligation mediated PCR on mitochondrial DNA sequencing", CHINA OCCUPATIONAL MEDICINE, vol. 27, no. 3, 30 June 2000 (2000-06-30), pages 16 - 18
LIU, LI ET AL.: "Analysis of the frequency and hot spot of mitochondrial DNA oxidative damage by ligation mediated PCR", CHINESE JOURNAL OF INDUSTRIAL HYGIENE AND OCCUPATIONAL DISEASES, vol. 18, no. 3, 30 June 2000 (2000-06-30), pages 151 - 154
O' MALLEY, R.C. ET AL.: "An adapter ligation-mediated PCR method for high-throughput mapping of T-DNA inserts in the Arabidopsis genome", NATURE PROTOCOLS, vol. 2, no. 11, 8 November 2007 (2007-11-08), pages 2910 - 2917, XP055249949
LIM, K.S. ET AL.: "Quantitative Mitochondrial DNA Mutation Analysis by Denaturing HPLC", CLINICAL CHEMISTRY, vol. 53, no. 6, 31 December 2007 (2007-12-31), pages 1046 - 1052, XP055439704
LIU, LI ET AL.: "Analysis of the frequency and hot spot of mitochondrial DNA oxidative damage by ligation mediated PCR", CHINESE JOURNAL OF INDUSTRIAL HYGIENE AND OCCUPATIONAL DISEASES, vol. 18, no. 3, 30 June 2000 (2000-06-30), pages 151 - 154
O' MALLEY, R.C. ET AL.: "An adapter ligation-mediated PCR method for high-throughput mapping of T-DNA inserts in the Arabidopsis genome", NATURE PROTOCOLS, vol. 2, no. 11, 8 November 2007 (2007-11-08), pages 2910 - 2917, XP055249949
LIM, K.S. ET AL.: "Quantitative Mitochondrial DNA Mutation Analysis by Denaturing HPLC", CLINICAL CHEMISTRY, vol. 53, no. 6, 31 December 2007 (2007-12-31), pages 1046 - 1052, XP055439704
Attorney, Agent or Firm:
ADVANCE CHINA IP LAW OFFICE (CN)
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