LAMP (loop-mediated isothermal amplification)-based fluorescence detection and CRISPR (clustered regularly interspaced short palindromic repeats)/Cas detection dual detection system and application

• Main Authors: XU DAN, HAN XU, WANG JUN, XIAO LINA, FANG ZHAOQING, GAO XIAOQING, ZHANG XINXIN
• Format: Patent
• Language: Chinese; English
• Published: 28.03.2023
• Subjects: BEER; BIOCHEMISTRY; CHEMISTRY; COMPOSITIONS OR TEST PAPERS THEREFOR; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL ORENZYMOLOGICAL PROCESSES; ENZYMOLOGY; 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 relates to an LAMP (loop-mediated isothermal amplification)-based fluorescence detection and CRISPR/Cas detection dual detection system and application thereof. Belongs to the technical field of nucleic acid detection. The invention relates to a double detection system for CRISPR/Cas and fluorescence detection based on LAMP (Loop-Mediated Isothermal Amplification). Comprising a target sequence amplification primer, a fluorescence report probe capable of identifying a target sequence, a fluorescence display agent for indicating whether a target product is generated in an isothermal amplification reaction, a reaction buffer solution, an enhancement buffer solution, an enzyme mixed solution containing DNA polymerase and CRISPR/Cas protease, and sgRNA capable of guiding the CRISPR/Cas protease to identify the target sequence. The kit has the characteristics of simplicity and convenience in operation, low use cost, high sensitivity and high specificity, and can simultaneously realize the following fu