Modified salting-out method: High-yield, high-quality genomic DNA extraction from whole blood using laundry detergent

被引:115
|
作者
Nasiri, H [1 ]
Forouzandeh, M [1 ]
Rasaee, MJ [1 ]
Rahbarizadeh, F [1 ]
机构
[1] Tarbiat Modarres Univ, Dept Med Biotechnol, Sch Med Sci, Tehran, Iran
关键词
DNA extraction; salting-out; polymerase chain reaction; genomic DNA; laundry detergent;
D O I
10.1002/jcla.20083
中图分类号
R446 [实验室诊断]; R-33 [实验医学、医学实验];
学科分类号
1001 ;
摘要
Different approaches have been used to extract DNA from whole blood. In most of these methods enzymes (such as proteinase K and RNAse A) or toxic organic solvents (such as phenol or guanidine isothiocyanate) are used. Since these enzymes are expensive, and most of the materials that are used routinely are toxic, it is desirable to apply an efficient DNA extraction procedure that does not require the use of such materials. In this study, genomic DNA was extracted by the salting-out method, but instead of using an analytical-grade enzyme and chemical detergents, as normally used for DNA isolation, a common laundry powder was used. Different concentrations of the powder were tested, and proteins were precipitated by NaCl-saturated distilled water. Finally, DNA precipitation was performed with the use of 96% ethanol. From the results, we conclude that the optimum concentration of laundry powder for the highest yield and purity isolated DNA is 30 mg/mL. The was optimized, and a final protocol suggested. Following the same DNA was extracted from 100 blood samples, and their amounts were found to >50 mu g/mL of whole blood. The integrity the DNA fragments was confirmed agarose gel electrophoresis. the extracted DNA was used as a for PCR reaction. The results obtained PCR showed that the final solutions extracted DNA did not contain any material for the enzyme used in the reaction, and indicated that the DNA was of good quality. These show that this method is simple, fast, and cost-effective, and can be used medical la- boratories and research centres.
引用
收藏
页码:229 / 232
页数:4
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