DNA: Methods for Preparation

Michael Mingfu Ling, Spectral Diagnostics Inc., Toronto, Canada

Published online: April 2001

DOI: 10.1038/npg.els.0002606

The isolation of DNA is required for genetic analysis and genetic engineering. There are primarily two types of method for isolating genomic DNA: (i) proteinase K digestion in the presence of SDS and EDTA at temperatures between 50 and 60°C; (ii) the use of strong chaotropic reagents such as guanidinium chloride. The trend is towards methods that are amenable to high-throughput screening and automation.

Keywords: DNA; genomic; isolation; genetic analysis; genetic engineering

 References
    Bowtell DD (1987) Rapid isolation of eukaryotic DNA. Analytical Biochemistry 162: 463–465.
    Cheng S, Chen Y, Monforte JA, Higuchi R and Van Houten B (1995) Template integrity is essential of PCR amplification of 20–30 kb sequences from genomic DNA. PCR Methods and Applications 4: 294–298.
    Chomczynski P and Sacchi N (1987) Single step method of RNA extraction by acid guanidinium thiocyanate–phenol–chloroform extraction. Analytical Biochemistry 162: 156–159.
    Gross-Bellard M, Oudet P and Chambon P (1973) Isolation of high-molecular-weight DNA from mammalian cells. European Journal of Biochemistry 36: 32–38.
    book Sambrook J, Fritsch EF and Maniatis T (1989) Molecular Cloning: A Laboratory Manual. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory.
 Further Reading
    Bignell GR, Miller AR and Evans IH (1996) Isolation of mitochondrial DNA. Methods in Molecular Biology 53: 109–116.
    Cooper A and Wayne R (1998) New uses for old DNA. Current Opinion in Biotechnology 9: 49–53.
    book Dieffenbach CW and Dveksler GS (1995) PCR Primer. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory Press.
    Evans WE and Relling MV (1999) Pharmacogenomics: translating functional genomics into rational therapeutics. Science 286: 487–491.
    book Rapley R (2000) Nucleic Acid Protocols Handbook. Totowa, NJ: Humana Press.

Related Sub-Topics:

Techniques and Tools in Clinical Genetics | DNA Structure and Function | Techniques and Tools in Molecular Biology
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Article Title: DNA: Methods for Preparation
 
How to Cite close
Ling, Michael Mingfu(Apr 2001) DNA: Methods for Preparation. In: eLS. John Wiley & Sons Ltd, Chichester. http://www.els.net [doi: 10.1038/npg.els.0002606]