咖啡ISSR与RAPD-PCR反应体系优化

以中粒种咖啡部分种质为试材，采用U25（55）均匀设计表，对影响ISSR和RAPD标记的模板DNA、Mg2+、dNTPs、TaqDNA聚合酶和引物浓度5个因素进行优化，分别建立适合于中粒种咖啡ISSR-PCR和RAPD-PCR标记的反应体系。结果表明：2种标记的反应体系相似，20 μL的反应体系中含DNA模板20 ng，Mg2+ 1.5 mmol/L，dNTPs 0.3 mmol/L，Taq酶1.25 U，引物0.6 μmol/L。利用所确立的体系对13份中粒种咖啡种质进行扩增，结果条带清晰明亮，多态性好

Optimization of RAPD and ISSR-PCR Reaction Systems for Coffee

The uniform design was used to optimize ISSR and RAPD reaction system for C.canephora,five influencing factors on ISSR and RAPD,including template DNA,Mg2+,dNTPs,TaqDNA polymerase and primer,concentration were tested by uniform design.The suitable ISSR-PCR and RAPD-PCR reaction systems for Coffea canephora were developed as follows: 20 μL PCR reaction volume containing 20 ng templates DNA,1.5 mmol/L Mg2+,0.3 mmol/L dNTPs,0.6μmol/L primer,1.25 U TaqDNA polymerase.Under this optimum system amplifications carried out on 13 genotypes of Coffea canephora produced clear polymorphic patterns.