Abstract: A method for constructing a consensus sequence from a sequence alignment. The consensus sequence may be used to generate molecular standards that may substitute for genomic DNA in various assays. Since a molecular standard cannot have unresolved bases, the method removes less informative sequences to resolve all positions in the alignment. Also includes several sequences from pathogenic waterborne species that were constructed according to the method.

Abstract: The present disclosure describes an adsorbent and exemplary protocols for extracting nucleic acids, such as DNA and RNA, from complex matrices, such as stool samples and water samples. The adsorbent is activated charcoal coated with a material such as polyvinylpyrrolidone, dextran, or coconut flours. The adsorbent may be used in microcentrifuge spin columns, where it may be present as a slurry in a storage solution. The sample may be prepared by vortexing in a buffer solution, centrifuging, adding a protease to the supernatant, and passing the supernatant through a microcentrifuge spin column containing coated activated charcoal. The key components, including buffer, protease, and spin columns, may be packaged in a kit.

Abstract: A method for constructing a consensus sequence from a sequence alignment. The consensus sequence may be used to generate molecular standards that may substitute for genomic DNA in various assays. Since a molecular standard cannot have unresolved bases, the method removes less informative sequences to resolve all positions in the alignment. Also includes several sequences from pathogenic waterborne species that were constructed according to the method.

Abstract: The present disclosure describes an adsorbent and exemplary protocols for extracting nucleic acids, such as DNA and RNA, from complex matrices, such as stool samples and water samples. The adsorbent is activated charcoal coated with a material such as polyvinylpyrrolidone, dextran, or coconut flours. The adsorbent may be used in microcentrifuge spin columns, where it may be present as a slurry in a storage solution. The sample may be prepared by vortexing in a buffer solution, centrifuging, adding a protease to the supernatant, and passing the supernatant through a microcentrifuge spin column containing coated activated charcoal. The key components, including buffer, protease, and spin columns, may be packaged in a kit.

Abstract: Disclosed is a rapid, dual purpose, PCR-based method for identifying two or more pathogens, including Giardia and/or Cryptosporidium, in an extracted sample, such as stool or environmental (soil, water) isolates, in individual real-time PCR reactions. This method is of particular utility for clinical, veterinary, and environmental testing applications. The present methods are more sensitive than conventional ELISA or IFA-based methods of detection. An internal control (IC) for use in the PCR based nucleic acid detection method is also disclosed.