Of a DNA extraction yield improved organizational methods.docVIP

 PAGE \* MERGEFORMAT 9 Of a DNA extraction yield improved organizational methods [Abstract] Objective conventional phenol - chloroform extraction method to extract organizational DNA, by extending the water-bath time to improve the yield of extracted DNA. Methods to set each group a different water bath time, compare the yield of DNA referred to. The results with phenol - chloroform extract of liver DNA, along with the extension of water bath time (2.5 ~ 7.5h), DNA extraction yield from each 100mg of liver tissue obtained 341.4μ g to 701.9μ g. Conclusion This method is suitable for use phenol - chloroform and a small number of organizations needed to extract the relatively large number of DNA experiments, for further experimental study preparation. [Keywords:] phenol - chloroform; DNA yield Genomic DNA in molecular biology research widely. Phenol - chloroform method of DNA isolated from tissues from the last century, application of the eighties, with its simple, effective and has been in the ascendant. This study was designed on the phenol - chloroform extract genomic DNA, the process of organizational improvement, a small amount of tissue extracted from a relatively large number of DNA, for the next experimental study [1]. 1 Materials and methods 1.1 Material obtained for each sample SD rat liver 0.1g, a total of 24 samples so as to sample a group of eight, each water bath at different times, Ⅰ group bath 2.5h; Ⅱ group of water-bath 5h; Ⅲ group bath 7.5h. 1.2 Main reagent cell lysate: 30mM Tris-HCl, 0.1mM EDTA, 0.1 mM NaCl, pH 8.0; 10mg/ml proteinase K, SDS, saturated phenol, chloroform, isoamyl alcohol, anhydrous alcohol, 1mol / L TE buffer fluid. 1.3 Methods 1.3.1 phenol - chloroform extraction Genomic DNA [2] (1) taking 100mg of liver plus 1ml cell lysis solution, homogenized with glass homogenizer; (2) homogenate poured into 2ml plastic centrifuge tube, add 20μ l protease K, coupled with SDS to a final concentration of 1%, upside d