利用正交设计优选菜心ISSR-PCR反应体系(英文)

Orthogonal design for optimal ISSR-PCR system in flowering Chinese cabbage

利用正交设计L16(44)探讨引物、Taq聚合酶、dNTPs、Mg2+对菜心ISSR-PCR反应的影响.得到适合菜心IS-SR-PCR反应的最佳体系:在25μL反应体系中含2.5μL 10×buffer,2.0 mmol/L Mg2+,0.5UTaqDNA聚合酶,0.24 mmol/L dNTPs,0.5μmol/L引物,60 ng DNA模板.

Factors which affected the ISSR-PCR reaction of flowering Chinese cabbage （Brassica campestris L. ssp. Chinensis Var. utilis Tssen. et Lee. ） , such as concentrations of primer, Taq DNA polymerase, dNTPs and Mg^2＋ , was studied by L16（44） orthogonal test. The optimal ISSR-PCR system for flowering Chinese cabbage was determined. In a total volume of 25 ul, containing 2.5 uL 10 × buffer, 2.0 mmol/L Mg^2 ＋, 0.5 units of Taq DNA polymerase, 0.24 mmol/L dNTPs, 0.50 umol/L primer and 60 ng of template DNA, clear and reproducibe DNA fragments can be obtained.