title: Oldies (but goldies!): extracting DNA from cryopreserved allozyme homogenates.
abstract: In this work, we describe a method that allows the extraction of plant genomic DNA from frozen enzyme homogenates. To test if reliable DNA was recovered from allozyme extracts, we compared these results with that obtained from fresh frozen and dried material. Sequencing and microsatellite fragments were analyzed for samples from 5 plant families in order to check for chloroplast and nuclear DNA quality. High-molecular weight DNA was recovered from frozen homogenates, and it was found that the quality was independent of storage time, which varied from 1 to 9 years. High-quality PCR products along with sequences and microsatellite fragments were obtained for nearly all analyzed samples. Frozen protein homogenates from previous population studies could thus be used for further phylogeographical and phylogenetic analyses using DNA molecular tools. This simple method provides an opportunity to take advantage of stored cryopreserved samples.
authors: Laboratorio Ecotono, Centro Regional Universitario Bariloche, Universidad Nacional del Comahue, Quintral 1250 (8400) Bariloche, Río Negro, Argentina. marbetman@gmail.com
authors: Marina P
authors: MP
authors: Arbetman
authors: Andrea C
authors: AC
authors: Premoli
mesh_terms: D018070:Biological Specimen Banks; D002467:Cell Nucleus / Q000235:genetics; D002736:Chloroplasts / Q000235:genetics; D015925:Cryopreservation; D016172:DNA Fingerprinting / Q000379:methods*; D018744:DNA, Plant / Q000032:analysis; D023281:Genomics / Q000379:methods*; D007527:Isoenzymes / Q000235:genetics*; D018895:Microsatellite Repeats; D010802:Phylogeny; D058974:Phylogeography; D010944:Plants / Q000235:genetics*; D016133:Polymerase Chain Reaction
chemical_list: D018744:DNA, Plant; D007527:Isoenzymes