悬铃木SSR反应体系建立及引物筛选

ESTABLISHMENT OF PLATANUS SSR MARKER REACTION SYSTEM AND SELECTION OF PRIMERS

以三球悬铃木组培苗为试验材料,对影响SSR-PCR扩增效果的退火温度、dNTP、引物浓度、Taq酶量及悬铃木DNA等因素的筛选,建立适宜的悬铃木SSR-PCR分子标记反应体系。结果表明,在20μL-1体积中,50℃退火温度、4 ng.μL-1模板DNA、0.2 mmol.L-1 dNTP、0.05 U.μL-1 Taq酶量和0.6μmol.L-1引物浓度是悬铃木的最适SSR反应体系。此外,利用该体系筛选出了54对适合悬铃木SSR扩增的引物。

The SSR reaction system of Platanus orientalis was established with its regenerated plantlets in vitro through optimizing some factors including annealing temperatures,Taq polymerase,the concentrations of dNTP,primer and template DNA.The results indicated that the optimized SSR reaction system for the plant was 50℃ anrealing temperature,4 ng · μL-1 template DNA,0.2 mmol · L-1dNTP,0.05 U · μL-1Taq enzyme and 0.6 μmol · L-1 primer.Moreover,54 pairs of primers were suitable and selected with this reaction system.