Genomic DNA Isolation Of Scale Insect And Microsatellite Primers Polymorphism PCR

The study examined the genomic DNA extraction from the 5 scale insect species and the polymorphisms PCR of the 5 species using the microsatellite primers. The 5 scale insect species was Ceroplastes ceriferus, Ceroplastes japonicus, Didesmococcus koreanus, Eulecanium gigantean and Phenacoccus fraxinus. The study is to explore an effective molecular genetic marker which will be useful in taxonomy and systematics. The results are as following:(1) The NaCl (sodium chloride) method is a good method for extracting the genomic DNA of the scale insects under the laboratory conditions. Through the tests about extracting the genomic DNA from different forms including nymph female imago, male imago and eggs, and samples which were processed and preserved in alcohol soaking, dried in oven, freezing and drying in the shade, the results showed that the method can be used in the genomic DNA extraction from the nymph female imago and eggs, also did the same to the alcohol preserved, dried in oven and freezing samples.(2) IdentifyingUnder the same action condition, we built a stable and well repeated PCR system using the SSR primers.Through the gradient temperature test, we got the fittest annealing temperature of each microsatellite primer: the 33.15 core sequence primer's was 54℃ and the (CAC)₅ primers' was 57℃. At any temperature test for the (GACA) ₄, we got no PCR strips, and we identified the (GACA) ₄ have no strips matching with the scale insects.The optimized PCR system:DNA template: 4μL, the primers: 15ng, Taq DNA polymerase: 2U, dNTP: 5mM, Mg²⁺: (25mmol/L) 2.5μL, 10×beffer: 2.5μL and...