| To optimize genome DNA extraction method of Chinese mitten crabs, genome DNA was extracted from gills,
muscle, blood and bristles of Chinese mitten crabs using phenol -chloroform method and DNA extraction kit, and the
quality of these DNA samples was evaluated in four aspects: the purity, concentration, fragment completeness and
microsatellite marker PCR amplification effect. The results showed that, compared with the method of DNA extraction kit,
the DNA extracted by phenol-chloroform method was much more complete, had higher concentration, but lower purity.
Both the extraction methods showed that the purity of DNA in bristles and blood were higher than that in the gills and
muscle, and the descending order of DNA content was gill, muscle, bristles, blood, while the uncutting bristles were failed
to extract genomic DNA. Microsatellite PCR electrophoresis results showed that the DNA extracted from four different
tissues by two dissimilar DNA extraction methods could meet the requirements of the application of microsatellite markers.
In conclusion, the present study established a convenient noninvasive sampling method and a procedure of DNA extraction
that could promote genetic breeding and molecular marker research of E. sinensis. |
