Extraction of total cellular DNA from plants, algae and fungi

Abstract

Historically, extraction of usable nucleic acids from plants and fungi has been difficult, in some instances notoriously so. In general, success in DNA extraction is measured by DNA yield, condition (molecular weight and color), and utility (or ease of use with restriction enzymes, polymerases, ligases, etc.). While yield is important, especially when milligram and submilligram amounts of fossil, dried or mummified tissues are used, it is less important than it once was due to the advent of PCR (polymerase chain reaction) methods. The condition of the DNA is similarly not as crucial as it once was. The utility, however, of the DNA is the paramount consideration in molecular biology manipulations. In a direct comparison [14] the DNAs (and RNAs) produced by methods employing cetyltrimethylammonium bromide (CTAB) [4, 8, 12, 15-19, 21, 23] generally exhibited lower levels of enzyme inhibition than did those by other methods [1, 3, 5, 9, 11, 13]. While the yields were lower with CTAB than for some of the other methods, the yields were still adequate for most uses in molecular biology and the condition of the DNA was above average.