DNeasy Plant and DNeasy 96 Plant procedures.
DNeasy Plant and DNeasy 96 Plant procedures.
PCR analysis.
PCR analysis.
RAPD analysis.
RAPD analysis.
Pure DNA (20–25 kb) for restriction analysis.
Pure DNA (20–25 kb) for restriction analysis.
DNA purity from oak leaves and pine needles.
DNA purity from oak leaves and pine needles.
DNeasy Plant and DNeasy 96 Plant procedures.
PCR analysis. DNA (10 ng) from the indicated leaves or needles was amplified using universal primers for the noncoding intergenic spacer between the tRNA genes trnL (UAA) 5' exon and trnL (UAA) 3' exon of cpDNA (Taberlet, P. et al. (1991) Plant Mol. Biol. 17, 1105). M: 100 bp ladder.
RAPD analysis. DNA (50 ng) from leaves of the indicated in vitro propagated sunflower (Helianthus) species was amplified using a 10-base RAPD primer and separated on a 1.5% agarose gel. M: 100 bp ladder. (Data kindly provided by H. J. Henn, Institute of Agricultural Botany, University of Bonn, Germany.)
Pure DNA (20–25 kb) for restriction analysis. Agarose gel (0.8%) analysis of DNA isolated from the indicated leaves or needles with the DNeasy Plant Mini Kit was perfromed. Undigested DNA (0.5 µg, left) or 1 µg of EcoRI-digested DNA (right) were loaded in each pair of lanes. M: lambda-HindIII.
DNA purity from oak leaves and pine needles. Spectrophotometric scans (220–320 nm) of DNA isolated from pine needles using the method of Dellaporta, CTAB, or the DNeasy Plant Mini Kit. Pure DNA typically shows a symmetrical peak at 260 nm and a smooth profile. Polysaccharides and other secondary metabolites, often copurified with plant DNA isolated using traditional methods, can interfere with OD readings (A260/ A280), leading to errors in determination of concentration and purity.