COI gene sequence of species of coral reef benthos and species identification method utilizing COI gene sequence
A benthic organism and gene sequence technology, applied in the fields of taxonomy and molecular biology, can solve the problems of lack of COI sequence information and less sequence information
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[0062] 1. Extraction of DNA template.
[0063] Take about 100mg of the muscle tissue of the sample to be tested (a sample of the dominant species in the community or a species with a large number of individuals), cut it into pieces or grind it evenly, and put it into a 1.5mL centrifuge tube. Use the Marine Animal Tissue Genomic DNA Extraction Kit (or any other effective method for extracting DNA) and follow the steps in the instructions to extract the genomic DNA of the sample. The obtained DNA was dissolved in TE buffer or ultrapure water (the pH of the solution was adjusted to about 8.0 with NaOH), and stored in a -20°C refrigerator. The extracted DNA was checked by 1.5% agarose gel electrophoresis, ethidium bromide (EB) staining and observation under ultraviolet light.
[0064] 2. PCR amplification of the COI gene sequence of the sample to be tested. The PCR reaction system for the COI gene sequence is as follows: a total volume of 50 μL. Among them, 5 μL of 10×PCR buffer ...
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