Hyla tsinlingensis microsatellite sites, primers for microsatellite sites and application of microsatellite sites and primers
A technology of microsatellite sites and hyla, which is applied in the determination/inspection of microorganisms, biochemical equipment and methods, DNA/RNA fragments, etc., and can solve problems such as blanks
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[0008] The present invention will be described in detail below in conjunction with specific embodiments.
[0009] 1. DNA extraction
[0010] Genomic DNA of Hyla Qinling was extracted using muscle or blood tissue. Genomic DNA was extracted using SDS / proteinase K cleavage and phenol-chloroform extraction (Sambrook et al., 1999).
[0011] 2. Construction of microsatellite enrichment library of Hyla Qinling
[0012] Genomic DNA was extracted, and AFLP rapid isolation method (Fast Isolation by AFLP of Sequences Containing repeats, FIASCO) was used to construct a microsatellite enrichment library. The specific process was as follows: Take about 250ng of genomic DNA, digest it with MseI, and at the same time combine it with the AFLP receptor of MseI Connector (5'-TAC TCA GGA CTC AT-3' / 5'-GAC GAT GAG TCC TGA G-3') for connection. After diluting the digested product tenfold, use AFLP receptor-specific primers (5'-GAT GAG TCC TGA GTA AN-3', referred to as MseI-N) for PCR amplificatio...
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