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A rapid, high yield mini-prep method for isolation of total genomic DNA from A rapid, high yield mini-prep method for isolation of total genomic DNA from fungi. fungi.

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Eumycetozoans, the myxomycetes, protostelids, and dictyostelids, were first hypothesized to be a monophyletic group by L.S. Olive, who suggested that the primitive members of the group were similar to some of the extant protostelids. A review of morphological evidence supporting some aspects of this hypothesis is presented along with explicit explanations of the shortcomings of morphological data as tests of other aspects. For the hypothesis to be supported, modified, or rejected, data from other areas such as the sequences of the nuclear ribosomal small subunit genes (SSrDNA) will have to be used. Presently, sequences for this gene are known only from Physarum polycephalum and Dictyostelium discoideum. These two slime molds are treated as separate, deep clades in the grand eukaryote phylogenies derived from the sequences of SSrDNA. That is, each species represents an independent lineage that diverged early in the history of the eukaryotes. Insufficient taxon sampling may account for the molecular trees which suggest that the dictyostelids and myxomycetes are not members of a monophyletic group. We have begun to examine the SSrDNA sequence in the protostelid Protostelium mycophaga. Preliminary phylogenetic reconstructions using 11 eukaryotic outgroups suggest that the protostelids, myxomycetes, and dictyostelids are members of a single monophyletic group which may be most closely related to the Chromista. It is interesting that these results coincide with earlier phylogenetic hypotheses based on the morphological characters of these slime molds. Key words: dictyostelids, myxomycetes, protostelids, ribosomal DNA, slime molds.

This review focuses upon a critical step in forensic biology: detection and quantification of human DNA from biological samples. Determination of the quantity and quality of human DNA extracted from biological evidence is important for several reasons. Firstly, depending on the source and extraction method, the quality (purity and length), and quantity of the resultant DNA extract can vary greatly. This affects the downstream method as the quantity of input DNA and its relative length can determine which genotyping procedure to use-standard short-tandem repeat (STR) typing, mini-STR typing or mitochondrial DNA sequencing. Secondly, because it is important in forensic analysis to preserve as much of the evidence as possible for retesting, it is important to determine the total DNA amount available prior to utilizing any destructive analytical method. Lastly, results from initial quantitative and qualitative evaluations permit a more informed interpretation of downstream analytical results. Newer quantitative techniques involving real-time PCR can reveal the presence of degraded DNA and PCR inhibitors, that provide potential reasons for poor genotyping results and may indicate methods to use for downstream typing success. In general, the more information available, the easier it is to interpret and process the sample resulting in a higher likelihood of successful DNA typing. The history of the development of quantitative methods has involved two main goals-improving precision of the analysis and increasing the information content of the result. This review covers advances in forensic DNA quantification methods and recent developments in RNA quantification.

Slot blot hybridization of membrane-immobilized, single-stranded human DNA with the higher primate-specific alphoid probe D17Z1 is routinely used in forensic science to estimate the amount of DNA in biological samples. Typically, a chemiluminescent signal captured on film records the hybridization, and the quantity of the signal is related to the amount of immobilized DNA. Digital imaging using a cooled CCD camera offers an alternate non-film-based method for image acquisition with comparable sensitivity of detection, a greater dynamic range, enhanced capability of data interpretation, and often faster results than film. In addition, the data support the premise that more accurate and precise human DNA quantification should be obtained by not assuming a linear response of signal to known standards. Instead, quantity should be estimated using a second-order standard curve (R2 = 0.999). Finally, a CCD camera imaging system offers versatility for image capture of different signal sources and analysis of samples on a variety of support media.