Construction method and application of sequencing library

• Main Authors: ZHANG LIMIN, ZHOU ZHENZHEN, SHI HONGJIE, CHEN DAFEI, CHEN YU, FENG JIANLONG, ZHANG FEI, NI ZHIWEI, WU XIN, JIANG WEI, YE LI
• Format: Patent
• Language: Chinese; English
• Published: 13.11.2020
• Subjects: BEER; BIOCHEMISTRY; CHEMISTRY; COMBINATORIAL CHEMISTRY; COMPOSITIONS OR TEST PAPERS THEREFOR; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL ORENZYMOLOGICAL PROCESSES; ENZYMOLOGY; LIBRARIES, e.g. CHEMICAL LIBRARIES, IN SILICOLIBRARIES; MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEICACIDS OR MICROORGANISMS; METALLURGY; MICROBIOLOGY; MUTATION OR GENETIC ENGINEERING; PROCESSES OF PREPARING SUCH COMPOSITIONS; SPIRITS; VINEGAR; WINE

The invention provides a construction method and application of a sequencing library. The method comprises the following steps: (1) respectively transferring M samples to be sequenced into competent bacteria, and carrying out streak culture for one night; (2) respectively picking N bacterial colonies from competent bacteria of each to-be-sequenced sample, and carrying out independent culture; (3)mixing the bacterium solutions containing different samples to be sequenced to form N mixed bacterium solutions, each mixed bacterium solution containing M competent bacteria of the samples to be sequenced; (4) respectively extracting plasmids from the N mixed bacterium solutions, and carrying out linear enzyme digestion; and (5) adding different tag sequences into the N linear mixed plasmids, andmixing to obtain a sequencing library; wherein M and N are positive integers. According to the method, a step of mixed extraction and enzyme digestion is adopted, the monoclonal screening process isomitted while the sample si