| By the orthogonal experimental design L16 (4 5), five factors, including Mg2+, dNTPs, Taq DNA polymerase, primer concentration and DNA template concentrations in Nilaparvata lugens SRAP PCR reaction system, were optimized, and the optimal reaction system of the brown planthopper SRAP PCR was established. The results showed that a suitable SRAP PCR reaction system was constructed with 10μL volume containing 25 ng of template DNA, 0.3 μmol/L of each of the two primers, 150μmol/L deoxynucleotide triphosphates (dNTPs), 3 mmol/L MgCl2, 1 × PCR buffer, and 2 unit of Taq DNA polymerase. We used the females of biotypes 1 and the males of biotype 2 monomer hybridization population for the optimal reaction system. The results showed that the bands were clear, with rich polymorphism and co dominant bands. It showed that the reaction system is stable and reliable. |
