A non-organic and non-enzymatic extraction method gives higher yields of genomic DNA from whole-blood samples than do nine other methods tested
| Autor: | S. Bye, Margaret Crisp, Debomoy K. Lahiri, Marion E. Hodes, John I. Nurnberger |
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| Rok vydání: | 1992 |
| Předmět: |
Serine Endopeptidases
Biophysics Chromosome Mapping DNA Ribonuclease Pancreatic Biology Biochemistry DNA extraction Chemistry Techniques Analytical chemistry.chemical_compound genomic DNA Restriction enzyme chemistry Solvents Humans Agarose Lymphocytes Endopeptidase K Restriction fragment length polymorphism Southern blot Whole blood |
| Zdroj: | Journal of Biochemical and Biophysical Methods. 25:193-205 |
| ISSN: | 0165-022X |
| Popis: | We compared ten methods for extraction of DNA from whole blood. Nine methods require incubation with either enzymes or treatment of organic solvents or both. The 'Rapid Method' (RM) (Method 10) avoids the use of organic solvents (phenol/chloroform) and eliminates completely the use of proteinase K. Thus, the time and cost of DNA extraction are reduced significantly. This is accomplished by salting out and precipitation of the cellular proteins in saturated sodium chloride. This method takes less than an hour to completion, without compromising the yield or the quality of DNA. Using RM, we can make DNA from 0.1 ml of whole blood and as little as 0.5 ml of blood yields DNA sufficient to run a few Southern blots. The RM can also be applied to packed cells. The DNA is free of RNA, protein and degrading enzymes. The uncut DNA runs as a typical slow-migrating, high-molecular-weight and undegraded species in an agarose gel. The DNA is suitable for digestion by various restriction endonucleases. This procedure works equally well with fresh blood samples and with those that are stored at 4 degrees C and -70 degrees C. To our knowledge the RM reported here is the safest, fastest and most quantitative and economical method for preparation of DNA from whole blood and cells. |
| Databáze: | OpenAIRE |
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