A simple and efficient method for isolating high molecular weight DNA from frozen mammalian tissues

The authors describe a very simple procedure which may be used to extract very high molecular weight DNA suitable for PFG analysis from frozen mammalian tissue. One gram of frozen tissue was ground to a fine powder under liquid nitrogen with a mortal and pestle. After the addition of 6 ml of TES the...

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Bibliographic Details
Published inNucleic acids research Vol. 16; no. 12; p. 5698
Main Authors PETRUKHIN, K. E, BROUDE, N. E, SMITH, C. L
Format Journal Article
LanguageEnglish
Published Oxford Oxford University Press 24.06.1988
Subjects
agarose
Analytical, structural and metabolic biochemistry
Biological and medical sciences
Cell Fractionation - methods
DNA - analysis
DNA - isolation & purification
Dna, deoxyribonucleoproteins
Freezing
Fundamental and applied biological sciences. Psychology
Humans
Molecular Weight
Nucleic Acid Hybridization
Nucleic acids
Vertebrata
Mammalia
DNA
Isolation
Technique
Freezing
Molecular weight
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Summary:The authors describe a very simple procedure which may be used to extract very high molecular weight DNA suitable for PFG analysis from frozen mammalian tissue. One gram of frozen tissue was ground to a fine powder under liquid nitrogen with a mortal and pestle. After the addition of 6 ml of TES the sample was homogenized using a Dounce, then filtered through two sheets of gauze. Nuclei were prepared by treating the cells with 1% triton X100 at 8 degree C for 10 min. Cell and nuclei suspensions mixed with an equal volume of 1% low gelling agarose were used to prepare insert samples.
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ISSN:0305-1048
1362-4962
DOI:10.1093/nar/16.12.5698