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在RAPD反应中,有许多因素影响结果的稳定性和准确性,本文以花生为材料,对RAPD各种反应条件的优化组合进行了摸索。结果表明,花生RAPD的较为理想的反应体系如下,10μl反应体积中含:50mmol/LKCl,10mmol/LTris-HCl(pH9.0),0.1%Triton-100,20～40ng引物,0.2nmol/μl的dNTPs,1～2μg/μl的BSA,15nmol/μlMgCl2,0.5～1单位Taq酶(以上均为SangonLtd.产品),30～60ng左右花生基因组DNA。
In the RAPD reaction, there are many factors that affect the stability and accuracy of the results. In this paper, the optimized combination of RAPD reaction conditions was explored using peanut as material. The results showed that the optimal reaction system of RAPD for peanut was as follows: 10 mmol / L KCl, 10 mmol / L Tris-HCl, pH 9.0, 0.1% Triton-100 and 20-40 ng primers, 0.2 nmol / dNTPs, 1 to 2 μg / μl BSA, 15 nmol / μl MgCl 2, and 0.5 to 1 Taq enzyme (all are products of Sangon Ltd.), and about 30 to 60 ng of peanut genomic DNA.