人参根际土壤微生物PCR-DGGE反应体系优化

[目的]优选人参根际土壤微生物的PCR-DGGE反应体系。[方法]以种植1年的人参土壤为材料,利用PCR-DGGE技术,对反应体系中的几种重要参数不同梯度进行了优化研究,包括模板、dNTPs、引物及Mg2+的用量。[结果]最适宜的反应体系为:模板DNA浓度为0.8μg/μl,dNTPs浓度为0.2 mmol/L,引物浓度为0.4μmol/L,Mg2+浓度为1.5 mmol/L。[结论]该方法简便快捷,为进一步研究人参根际土壤微生物多样性奠定了基础。

Optimization of PCR-DGGE Reaction System on Rhizosphere Soil Microorganism for Panax ginseng C.A.Mey.

[Objective] To explore the PCR-DGGE reaction system on rhizosphere soil microorganism for Panax ginseng C.A.Mey..[Method] Some important factors for PCR-DGGE amplification system including the concentration of DNA template,Mg2+,dNTPs and primers were optimized with 1-year-old P.ginseng soil as materials.[Result] An optimal system(25 μl) was obtained as 0.8 μg/μl DNA template,0.2 mmol/L dNTPs,0.4 μmol/L primer,and 1.5 mmol/L Mg2+.[Conclusion] The method is simple and fast.This research can laid basis for further analyzing the genetic diversity of rhizosphere soil microorganism for P.ginseng.