3 articles published in JoVE
Isolation and Genome Analysis of Single Virions using 'Single Virus Genomics' Lisa Zeigler Allen1, Thomas Ishoey1, Mark A. Novotny1, Jeffrey S. McLean1, Roger S. Lasken1, Shannon J. Williamson1 1Department of Microbial and Environmental Genomics, The J. Craig Venter Institute Single Virus Genomics (SVG) is a method to isolate and amplify the genomes of single virons. Viral suspensions of a mixed assemblage are sorted using flow cytometry onto a microscope slide with discrete wells containing agarose, thereby capturing the virion and reducing genome shearing during downstream processing. Whole genome amplification is achieved using multiple displacement amplification (MDA) resulting in genomic material that is suitable for sequencing.
The Green Monster Process for the Generation of Yeast Strains Carrying Multiple Gene Deletions Yo Suzuki1, Jason Stam1, Mark Novotny2, Nozomu Yachie3, Roger S. Lasken2, Frederick P. Roth3,4 1Department of Synthetic Biology and Bioenergy, J. Craig Venter Institute, 2Department of Microbial and Environmental Genomics, J. Craig Venter Institute, 3Donnelly Centre & Department of Molecular Genetics, University of Toronto, 4Lunenfeld Research Institute, Mt Sinai Hospital The Green Monster method enables the rapid assembly of multiple deletions marked with a reporter gene encoding green fluorescent protein. This method is based on driving yeast strains through repeated cycles of sexual assortment of deletions and fluorescence-based enrichment of cells carrying more deletions.
Separation of Single-stranded DNA, Double-stranded DNA and RNA from an Environmental Viral Community Using Hydroxyapatite Chromatography Douglas W. Fadrosh1, Cynthia Andrews-Pfannkoch2, Shannon J. Williamson1 1Department of Microbial and Environmental Genomics, The J. Craig Venter Institute, 2Department of Synthetic Biology and Bioenergy, The J. Craig Venter Institute We describe an efficient method to separate single-stranded DNA, double-stranded DNA and RNA molecules from environmental viral communities. Nucleic acids are fractionated using hydroxyapatite chromatography with increasing concentrations of phosphate-containing buffers. This method permits the isolation of all viral nucleic acid types from environmental samples.