Influencing factors on Genomic DNA extraction from whole blood using modified salting-out method

【Objective】 To observe the effects of erythrocyte lysis time,DNA purification method and DNA drying time on genomic DNA extraction from whole blood using modified salting-out method.【Methods】 Different erythrocyte lysis time,DNA purification method and DNA drying time were used for genomic DNA extraction from whole blood using modified salting-out method.Then,the cell counts and percentage of dead cells were observed by flow cytometry after erythrocyte lysis.The DNA concentration and OD260 / OD280 were observed by ultraviolet spectrophotometer.Finally,PCR products were detected by agarose gel electrophoresis.【Results】 The percentage of dead cells increased but the cell counts,the DNA production and OD260 / OD280 had no significant change when erythrocyte lysis time used from 10 min to 120 min.The DNA production and OD260 / OD280 purified by isopropanol were greater than purified by silica membrane column.DNA drying time from 30 min to 120 min had no influence on DNA production,OD260 / OD280 and PCR.【Conclusions】 When modified salting-out method was used for genomic DNA extraction from whole blood,erythrocyte lysis time could be increased to 120 min,DNA drying time could be decreased to 30 min,and DNA purified by isopropanol was better than purified by silica membrane column.