Establishment and Optimization of ISSR Reaction System of Schima superba
In order to study the genetic diversity of Schima superba with ISSR technique, the factors which affected the ISSR amplification such as template DNA dosage, primers dosage, bovine serum albumin concentration, unit of Taq DNA polymerase, dNTP concentration, Mg2+ concentration and annealing temperature were selected and optimized. The results showed that the conditions being suitable for ISSR-PCR of Schima superba were as follows: 1×Taq buffer(10mmol/LIris-HCI, pH9. 0, 50 mmol/L KC1 and 0. 1 % Triton X-100), 0. 75 U Taq DNA polymerase, 1. 5 mmol/L MgCl2, 0. 2 mol/L 4 × dNTP, 6 pmol primers, 2 mg/ml BSA and 10 ng template DNA. The suitable annealing temperature of Schima superba in the ISSR-PCR reaction system was 56. 3 ℃.
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