Isolation of DNA from plant cells by gel filtration on agarose

✍ Scribed by P.F. Lurquin; G. Tshitenge; G. Delaunoit; L. Ledoux

• Publisher: Elsevier Science
• Year: 1975
• Tongue: English
• Weight: 568 KB
• Volume: 65
• Category: Article
• ISSN: 0003-2697
• DOI: 10.1016/0003-2697(75)90483-2

✦ Synopsis

A method is described which allows the obtention of total DNA from cells of Chlamydomonas reinhardi in 1 day.

It is based on the gel filtration on agarose of crude lysates incubated for 1 hr with pronase and subsequently brought to 2 M NaCl in order to dissociate nucleoprotein complexes. The DNA obtained in this way contains no detectable pigments nor RNA and is contaminated by less than 2% protein. The buoyant densities determined by CsCl density gradient centrifugation are 1.724 * 0.002 and 1.695 ? 0.002 g/cm3 for nuclear and chloroplast DNA, respectively. A slight modification of the method makes it applicable to the extraction of DNA from Vigna mungo (Phaseolus aureus) whose buoyant density in a CsCl gradient is found to be 1.693 ? 0.002 g/cm3.

The molecular weight of DNA extracted from both systems can be grossly estimated to be around 3-10 x 106.