Construction method and application of genome-wide methylation high-throughput sequencing library and

• Main Authors: GAO FEI, WANG JUN, WANG JUNWEN, XIA YUDONG
• Format: Patent
• Language: Chinese; English
• Published: 08.05.2013
• Subjects: APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; BEER; BIOCHEMISTRY; CHEMISTRY; COMBINATORIAL CHEMISTRY; COMPOSITIONS OR TEST PAPERS THEREFOR; COMPOSITIONS THEREOF; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL ORENZYMOLOGICAL PROCESSES; CULTURE MEDIA; ENZYMOLOGY; LIBRARIES, e.g. CHEMICAL LIBRARIES, IN SILICOLIBRARIES; MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEICACIDS OR MICROORGANISMS; METALLURGY; MICROBIOLOGY; MICROORGANISMS OR ENZYMES; MUTATION OR GENETIC ENGINEERING; PROCESSES OF PREPARING SUCH COMPOSITIONS; PROPAGATING, PRESERVING OR MAINTAINING MICROORGANISMS; SPIRITS; VINEGAR; WINE

The invention provides a construction method and application of a genome-wide methylation high-throughput sequencing library. The construction method of the genome-wide methylation high-throughput sequencing library comprises steps of: conducting digestion on a genome DNA with Msp I and a second restriction endonuclease; conducting end repair on DNA fragments; adding a basic group A on a 3'terminal of the DNA fragment subjected to end repair; connecting a DNA fragment with a cohesive end A to a methylation joint; conducting fragment selection on the connect products with the methylation joint, in order to obtain a target fragment; subjecting the target fragment to a bisulfite treatment, in order to convert unmethylated cytosine in the target fragment to uracil; subjecting the converted target fragment to PCR amplification; and separating and purifying the amplification products, wherein the amplification products form the genome-wide methylation high-throughput sequencing library. The construction method and