Extraction of high-quality genomic DNA from latex-containing plants.
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 cetyltrimethylammonium bromide 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 25 degrees C isopropanol precipitation step. The purified DNA has excellent spectral qualities, is efficiently digested by restriction endonucleases, and is suitable for long-fragment PCR amplification.
Michiels
An
A
Laboratory of Plant Development, Institute of Botany and Microbiology, K. U. Leuven, Belgium. an.michiels@bio.kuleuven.ac.be
Van den Ende
Wim
W
Tucker
Mark
M
Van Riet
Liesbet
L
Van Laere
André
A
D019659:Asteraceae / Q000235:genetics*; D015139:Blotting, Southern; D004262:DNA Restriction Enzymes / Q000378:metabolism; D018744:DNA, Plant / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D010946:Plants, Medicinal / Q000235:genetics; D016133:Polymerase Chain Reaction; D013696:Temperature; D013997:Time Factors
D018744:DNA, Plant; D004262:DNA Restriction Enzymes