Monitoring method for natural reproduction of fish based on environmental DNA technology
A technology of natural reproduction and technology, applied in the field of molecular ecology, can solve the problems of high operation difficulty, high labor intensity, and different hydrological conditions, and achieve the effect of simplified operation steps and high detection sensitivity
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Embodiment 1
[0033] The natural reproduction of domestic fish during the first operation of the Three Gorges Reservoir in 2016 (June 8-12, 2016). 1. Sample collection
[0034] According to many years of traditional survey results, six sections were set up to collect water samples within the 36-kilometer section from Yichang Miaozui Wharf to Houjiangtuo. Take 2L of mixed water samples from water layers of different depths and store them in an incubator at 4°C. The specific geographical locations of the sampling points are shown in Table 1. 2. DNA extraction
[0035] Vacuum filter the above water sample through a fat-soluble filter membrane with a pore size of 0.22 μm within 24 hours. According to multiple pre-experimental comparisons, one filter membrane should be used for each liter of water, so put the filtered two filter membranes into Store in a 50ml centrifuge tube at -20°C for later use.
[0036] Using a professional water sample DNA extraction kit the Water DNA Kit (Omega) was ...
Embodiment 2
[0058] Embodiment 2: The natural breeding situation of domestic fish when the Three Gorges Reservoir was dispatched for the second time in 2016 (June 19th-22nd, 2016), the method steps are the same as in Example 1, and Table 3 is 6 sections and 18 sampling points are continuous For the DNA copy number of domestic fish obtained by ddPCR quantitative analysis in 4 days, the daily average total concentration can be found in image 3 , the concentration of each section see Figure 4 .
[0059] Table 3. DNA copy numbers of domestic fish detected during the second dispatch
[0060]
[0061] According to the experimental results of ddPCR, the copy number of the target species contained in each sample is known, and the daily average total DNA concentration of domestic fish eDNA in 6 sections of the monitoring period is compared. It can be known that the domestic fish environment on the first day of the second ecological dispatch DNA content is maximum. Comparing the DNA concentr...
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