Real-time constant-temperature index amplification method of tiny RNAs

• Main Authors: FU WEILING, XIA HAN, HUANG JUNFU, HUANG QING
• Format: Patent
• Language: English
• Published: 16.12.2015
• 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 discloses a real-time constant-temperature index amplification method of tiny RNA target molecules. In the method, a specific upstream primer and a specific downstream primer of the target molecules are provided, anchoring sequence regions of the specific upstream primer and the specific downstream primer contain gap agent identifying sequence positive-sense strand sequences, recognition sequence regions are respectively complementary with 3'-end sequences of the target molecules and the complementary strands thereof, the upstream primer and/or the downstream primer contain(s) interactive marking systems, so that the index amplification of the target molecules under the steady temperature condition is realized, specific detectable signals of the target molecules are generated through the interactive marking systems contained by the primers, and thus qualitative and/or quantitative detection for one or more kinds of target molecules is realized in one detection unit. With the adoption of the method, the design difficulty of the amplification detection method caused by the inherent attributes of the tiny RNAs is overcome, a constant-temperature amplification system based on the oligonucleotide primers is provided, the index amplification of the target molecules is realized under the single steady temperature condition, and multiple kinds of target molecules are simultaneously detected in one detection system.