Chinese giant salamander group dividing method based on mitochondrial DNA sequence
A DNA sequence and group division technology, applied in the field of biological identification, can solve the problems of difficult group division standards, no group markers, unstable body surface patterns, etc., and achieve the effects of easy operation, low experimental operation cost, and easy mastery
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[0041] (1) Sample collection and DNA extraction
[0042] Chinese giant salamander individuals were collected from Xing'an in Guangxi (GXXA), Longshan in Hunan (HNLS), Jing'an in Jiangxi (JXJA), Huangshan in Anhui (AHHS), Laifeng in Hubei (HBLF), and Lishui in Zhejiang (ZJLS). The part or tissue collected can be a small amount of fin rays, blood, muscle, fresh molted skin, or oral epithelial cells scraped with a sterilized cotton swab, etc. Genomic DNA was extracted using DNA extraction kit or phenol-chloroform method. Use a spectrophotometer to measure the purity and content of the DNA, and store it at -20°C for future use. The DNA concentration was adjusted to 10-100ng / μL when used.
[0043] (2) Primer synthesis
[0044] The following primers were synthesized at Shanghai Sangon Biotechnology Co., Ltd.:
[0045] L14764: 5′-ATGAAACAGGGTCAAAGCAATCCAAC-3′
[0046] H16062: 5'-AGATGAGGGCAGACTCAGTTATG-3'.
[0047]Dissolve and dilute the primers to a concentration of 10 μmo / L w...
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