Specific high-throughput sequencing method for single-stranded DNA based on 5 'connection

• Main Authors: LIU DEPEI, XIE YAN, MA LANZHI
• Format: Patent
• Language: Chinese; English
• Published: 15.03.2024
• Subjects: 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 belongs to the technical field of biology, and particularly relates to a specific high-throughput sequencing method for single-stranded DNA based on 5'connection. The method specifically comprises a method for preparing a single-stranded DNA sequencing library based on 5 '-terminal connection. The method comprises the following steps for treating a sample to be detected: step 1) supplementing the 5'-terminal of dsDNA with DNA polymerase without exonuclease activity; the method comprises the following steps of (1) carrying out 3'terminal tailing, (2) carrying out 3 'terminal tailing, (3) connecting a 5'-stem-loop joint subjected to renaturation treatment, (4) carrying out amplification on a connection product, and (5) carrying out amplification on the connection product to form a single-stranded DNA sequencing library, wherein the structure of the 5 '-stem-loop joint is as follows: a protruding terminal, a random base region, a first stem region, a loop region and a second stem region, and the fi