|Michiels, An - K.U. LEUVEN, BELGIUM|
|Van Den Ende, Wim - K.U. LEUVEN, BELGIUM|
|Van Riet, Liesbet - K.U. LEUVEN, BELGIUM|
|Van Laere, Andre' - K.U. LEUVEN, BELGIUM|
Submitted to: Analytical Biochemistry
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: January 1, 2003
Publication Date: N/A
Interpretive Summary: Extraction of good quality DNA from plants that produce latex-like substances (sticky, milky-white secretion) has been a problem for many laboratories. The quintessential latex producing plant is that used to make rubber; however, many more common plants also produce a latex-like secretion when injured, e.g., chicory, endive, lettuce and dandelion. Currently used standard procedures were adapted and optimized for extraction of high quality DNA from latex producing plants. This is a significant accomplishment that will aid many scientists in their molecular biology research on these latex producing plant species.
Technical Abstract: The isolation of intact, high molecular mass genomic DNA is essential for many molecular biology applications including long PCR, endonuclease restriction digestion, Southern blot analysis and genomic library construction. Many protocols are available for the extraction of DNA from plant material. However, for latex-containing Asteraceae (Cichorioideae) species, standard protocols and commercially available kits do not produce efficient yields of high quality amplifiable DNA. A CTAB protocol has been optimized for isolation of genomic DNA from latex-containing plants. Key steps in the modified protocol are the use of etiolated leaf tissue for extraction and an overnight 25ºC isopropanol precipitation step. The purified DNA has excellent spectral qualities, is efficiently digested by restriction endonucleases and is suitable for long fragment PCR amplification.