title: An efficient DNA extraction method for desert Calligonum species.
abstract: Genetic conservation programs in arid environments rely on molecular methods for diversity assessments. DNA-based molecular profiling will aid in conservation and protection of species from genetic erosion. Obtaining intact genomic DNA from Calligonum species, of sufficiently high-quality that is readily amplifiable using PCR, is challenging because of the presence of the exceptionally large amount of oxidized polyphenolic compounds, polysaccharides, and other secondary metabolites. The present method involves a modification of the available CTAB method employing higher concentrations of NaCl and CTAB, and incorporating PEG 6000 (1%) and glucose. The yield of DNA was 60-670 μg g(-1) of fresh tissue. The protocol has been tested with two species from the arid region. The DNA isolated was successfully amplified by two ITS primer pairs. PCR-RFLP analysis of the ITS1-5.8S-ITS2 region among and within Calligonum species followed by sequencing is under way.
authors.affiliation: Laboratoire d'Ecologie Pastorale, Institut des Régions Arides, 4119 Médenine, Tunisia. raoudhamabdellaoui@yahoo.com
authors.forename: Raoudha
authors.initials: R
authors.lastname: Abdellaoui
authors.forename: Hassen
authors.initials: H
authors.lastname: Gouja
authors.forename: Amel
authors.initials: A
authors.lastname: Sayah
authors.forename: Mohamed
authors.initials: M
authors.lastname: Neffati
mesh_terms: D017931:DNA Primers / Q000737:chemistry; D018744:DNA, Plant / Q000235:genetics / Q000302:isolation & purification*; D019669:Embryophyta / Q000737:chemistry / Q000235:genetics*; D018745:Genome, Plant; D016133:Polymerase Chain Reaction / Q000379:methods; D012150:Polymorphism, Restriction Fragment Length; D013045:Species Specificity
chemical_list: D017931:DNA Primers; D018744:DNA, Plant