扩增条件对茶类植物RAPD带的影响

IMPACTS OF RAPD AMPLIFICATION CONDITIONS ON RAPD BANDS FOR CAMELLIA

摘要:

采用梯度分析的方法试验了模板DNA、引物、镁离子、dNTP和Taq酶的浓度对茶类植物进行RAPD分析中DNA扩增结果的影响。实验表明这些条件的变化对扩增出来的RAPD带的数目和强弱会产生影响。经过比较分析，筛选出对于茶类植物进行ＲＡＰＤ分析较理想的扩增条件：2.0mmol/L MgCl_{2，200 μmol/L dNTP，15 ng引物/20 μl反应体积，4 ng模板DNA/μl反应体积，1 U Taq酶/20 μl反应体积。}

Abstract:

The impacts of the concentrations of template DNA, primer, magnesium, dNTPand Taq DNA polymerase on RAPD amplification for Camellia were tested by means ofladder analysis. The results showed that RAPD band patterns were changed observablywith the variation of concentrations of template DNA or primer or Taq DNA polymerase,and there was a positive interrelation between the both within a limited range, but littleimpact was generated by the concentrations of 1.0-4.0 mmol/L MgCl₂ or 100-400 μmol/LdNTP on RAPD band patterns The optimal amplification conditions for Camellia were chosen as follows：4 ng template DNA／μl reaction volume，15 ng primer／20 μl reaction volume，2.0 mmol／L MgCl₂，200／μmol／L dNTP．1 U Taq DNA polymerse／20 μl reaction volume.