The mucopolysaccharide capsule of Cryptococcus neoformans and other pathogenic yeasts prevent the extraction of DNA from these important zoonotic agents. We report that the use of a lysis buffer containing a high concentration of urea is an easy, efficient and time-saving technique to obtain high yields of good-quality DNA for molecular diagnosis. The use of urea also prevents the degradation of DNA during storage of samples at room temperature for up to 6 months.
Beginning with a description of the conventional Giemsa-stained karyotype of the tench (Tinea tinea L.), the structure and variability of the chromosomal heterochromatic regions in this cyprinid species were analyzed by means of C-, silver-, and restriction endonuclease banding. Silver staining revealed active nucleolus organizer regions (NORs) on the secondary constriction of chromosome pair 3. Constitutive heterochromatin was associated with NOR regions detected by C-banding. Restriction endonuclease digestion with AluI, TaqI, and HaeIII induced specific banding patterns that allowed identification of homologous chromosome pairs and revealed features about the sequence composition of several chromosomal heterochromatic regions and of the NOR-associated heterochromatin.
Lesser kestrel, Falco naumanni, is a colonial and migratory species breeding in part of the Mediterranean Basin and part of central Asia and north-east of China and Mongolia. This species is catalogued in IUCN red list category as vulnerable. Twenty microsatellite loci were selected from libraries enriched in AC or AG tandem repeats and specific PCR were devised from their flanking sequences. Most microsatellites (14) were found polymorphic among 30 individuals of F. naumanni representing 20 reproduction areas of the species in the region of Extremadura, Spain. Polymorphisms were detected by size variation of the amplified loci, which allele number and observed heterozygosity ranged from 3 to 20 and from 0.300 to 0.933, respectively. Cross-species amplification showed that 13 of selected loci were also found polymorphic in common kestrel species (Falco tinnunculus). Novel polymorphic microsatellites will serve to conservation studies in lesser kestrel.
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