Whole Blood Rapid Extraction Of Genomic Dna And Its Applications

Objective: To develop a simple, rapid, reliable and inexpensive method for extracting genomic DNA from whole blood. A improved Guanidinium thiocyanate-silica gel isolation procedure based on solid phase vector method was establishedMethods: Guanidinium thiocyanate-silica gel method was established .Firstly, cell nuclei are prepared by lysis of cytoplasmic membrance. DNA within the nuclear pellet is dispersed with Guanidinium thiocyanate and selectively bound to silica gel.Then the silica gel was washed in wash buffer. Finally, the DNA is recovered by elution in TE. Six factors were used to evaluate the Guanidim'um thiocyanate-silica gel method:①DNA yield and DNA purity ②DNA integration③Assessment of RNA removal ④reproducibility ⑤PCR ⑥restriction enzyme digestion. And it was compared with conventional phenol-chloroform extraction method.Result: The Guanidinium thiocyanate-silica gel isolation procedure resulted in higher yield and intact DNA.The DNA purity obtained by the Guanidinium thiocyanate-silica gel method was comparable to that obtained by phenol-chloroform extraction method. The DNA yield and purity CV of Guanidinium thiocyanate-silica gel method was 6.02% and 6.74% respectively.The DNAs extracted by Guanidinium thiocyanate-silica gel method have been successfully used for PCR and restriction enzyme digestion,which were completely concordart with that extracted by phenol-chloroform method.Some factors such as temperature, pH and concentration of Guanidinium thiocyanate were of important for efficiency of Guanidinium thiocyanate-silica gel methodConclusion: The Guanidinium thiocyanate-silica gel isolation procedure is simple, rapid, reliable method for extracting high-quality DNA in high yield,which is suitable for PCR, restriction enzyme digestion and the other experiments.