Extraction Of Medicinal Plant Genomic DNA And Optimizing Of RAPD Reaction System In Dendrobium Candidum Wall Ex Lindl.

As the life science developing, the molecular biotechnology is increasingly permeated into medicinal botany damain. The way of Chinese drugs identification detecting gene polymorphic molecular markers is fully used, including RAPD which is simple, fast, effective. RAPD is widely applied for many researches, such as variety identification of plant medicinal materials, the species specificity study of origin plant, the adulterants authentication, the genuine study of the medicinal materials and so on.The high yields and purity genomic DNA is the basic in RAPD. 13 medicinal plant collected from emei mountain, Sichuan is used to explored a DNA extracted approach which is fast and effective and also can be used to many medicinal plant. After the step of KAc removeing denaturating protein in traditional DNA extracted way, it is added that using Phenol: Chloroform extracting the supernatant two or three times. And in the nucleic acid precipitating process, the nucleid acid floccule can be hooked fast preventing the polysaccharide and phenol from contamining the DNA. Sepharose electrophoresis shows that the quality of the DNA is good so that it can satisfy the following experiment level.The modified SDS method was used to extract DNA from Dendrobium candidum Wall ex Lindl.. The value of A₂₆₀/A₂₈₀ is between 1.8 and 2.0. Sepharose electrophoresis shows that its integrality is better. Using this DNA as template, the effect of the concentrations of dNTPs, MgCl₂, Primer and template was evaluated in RAPD-PCR amplification system. The results indicated that the optional concentrations of dNTPs, MgCl₂, Primer and Template is 0.25 mmol/L, 1.50 mmol/L, 0.32μmol/L, 0.40 ng/μl in 25μl PCR reaction system.