Method for realizing absolute quantification of blue-green algae at genus level based on next-generation sequencing technology

• Main Authors: ZHANG PEIPEI, SUN BEILI, ZHOU KERU
• Format: Patent
• Language: Chinese; English
• Published: 01.01.2021
• 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 method for realizing absolute quantification of blue-green algae at a genus level based on a next-generation sequencing technology, and relates to the related field of blue-green algae absolute quantification. Accurate absolute quantification of a blue-green algae population at the genus level is realized by adding an artificially synthesized DNA internal reference into asample and meanwhile correcting the copy number of 16S rRNA genes of each genus under cyanophyta, and the real change of the blue-green algae flora structure is revealed. The method comprises the following steps: S1, a P-Spikes sequence is synthesized and cloned to a plasmid pMA-T, and a plasmid is guided into Escherichia coli; S2, a known amount of P-Spikes is added into an environmental sampleto be subjected to DNA extraction and PCR amplification, agarose gel electrophoresis detection is completed through a reaction, and purification treatment is conducted through a PCR product purification kit; S3, a library is e