西瓜种植物RAPD分析影响因子的研究

Studies on Affecting Factors in the Analysisof the Plant of Clanatus

以西瓜种内的１２个品种（系）为模板ＤＮＡ，研究了影响ＲＡＰＤ扩增效果的因素。结果表明，模板ＤＮＡ、Ｔａｑ酶、随机引物、Ｍｇ２＋浓度以及ＰＣＲ反应程序在ＲＡＰＤ分析中都具有重要的作用。根据以上因子的研究结果，建立了适合西瓜种植物的ＲＡＰＤ分析的技术体系：１０μＬ反应液中，模板１０ｎｇ／１０μＬ，引物０５μｍｏｌ／Ｌ，ＴｒｉｓＨＣｌ（ｐＨ８３）５０ｍｍｏｌ／Ｌ，ＢＳＡ５００μｇ／ｍＬ，ＭｇＣｌ２２５ｍｍｏｌ／Ｌ，ＴａｑＤＮＡ０６Ｕ／１０μＬ，ｄＮＴＰｓ２００μｍｏｌ／Ｌ，１０％蔗糖４００和１ｍｍｏｌ／Ｌ甲酚红。

The studies of affectors affecting RAPD were carried out by the 12 species of CLanatus as template DNAThe results showed that the purification of template DNA,the concentrations of template DNA,Taq polymerase,random primer,Mg2+and the PCR programe played an important role in RAPD analysisBased on the researches above,a technical system for RAPD analysis in species of Clanatus was established:in 10 L reaction solution,containing 10 ng/10 L template DNA,05 mol/L random primer,50 mmol/L TrisHCl(pH83),500 g/mL BSA,25 mmol/L MgCl2,06 U/10 L Taq DNA,200 mol/L dNTPs