【摘要】 利用不同的引物、引物浓度、复性温度、ｄＮＴＰｓ浓度、Ｔａｑ酶含量等对棉花品种鄂荆１号、ＰｉｍａＳ－３进行ＲＡＰＤ分析。结果表明，高温复性有利于提高扩增带型的特异性。低Ｍｇ２＋浓度易导致小片断扩增，高Ｍｇ３＋浓度易产生非特异带型。ｄＮＴＰｓ浓度增加导致扩增带型的数目减少。双引物扩增带型与卓引物相比可以检测到更多的品种间遗传差异。根据上述分析，棉花ＲＡＰＤ分析反应物组成为：１Ｘ反应ｂｕｆｆｃｒ、１．８ｍｍｏｌ．Ｌ－１Ｍｇ２＋、０．１６ｍｍｏｌ·Ｌ－１ｄＮＴＰｓ、４０ｎｇＤＮＡ、０．６μｍｏｌ．Ｌ－１随机引物、Ｔａｑ酶１．５单位，加双蒸水至２５μｌ反应体系。更多还原

【Abstract】 RAPD analysis on cotton varities Ejing and PimaS-3 with different perimers,concentration of priers,Mg2+ and Taq polymerase annealing temperature,dNTPs,etc.were conducted.The results showed that the specificity of polymorphric DNA fragment wasimproved at high temperature.For the same primer,small fragments were amplified at lowMg2+ concentration and many unspecific fragments resulted from much too high Mg2+ con-centration.Less fragment were detected while increasing concentration of dNTPs.Morepolymorphric DNA fragments were observed by PCR with double primers than with single1.8mmol.L-1Mg2+,0.16mmol.L-1 dNTPs,40ngDNA,1.5Units Taq polymerase,1.6μmmol.L-1random primer,dd H2O was added to 25μl total volume.更多还原