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University of Georgia (UGA)

4 ARTICLES PUBLISHED IN JoVE

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Biology

Analyzing Gene Expression from Marine Microbial Communities using Environmental Transcriptomics
Rachel S. Poretsky 1, Scott Gifford 1, Johanna Rinta-Kanto 1, Maria Vila-Costa 1, Mary Ann Moran 1
1Department of Marine Sciences, University of Georgia (UGA)

We present a method for generating cDNA from environmental mRNA. In general, total RNA is first collected from the environment, rRNA is selectively removed, mRNA is selectively amplified, and cDNA synthesized from the enriched mRNA pool is sequenced. Recovered sequences can be annotated using standard bioinformatics techniques to identify the expressed genes.

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Biology

Processing the Loblolly Pine PtGen2 cDNA Microarray
W. Walter Lorenz 1, Yuan-Sheng Yu 1, Marta Simões 2, Jeffrey F. D. Dean 1
1Warnell School of Forestry and Natural Resources, University of Georgia (UGA), 2Instituto de Biologia Experimental e Tecnológica, Instituto Tecnologia Química e Biológica UNL, Av. da República

The cDNA microarray PtGen2 was developed for gene expression studies in loblolly pine, P. taeda, and other conifer species. Here, we show pre- and post-hybridization handling and washing techniques that can be used with this array to yield better consistency, reduced artifacts, and lower backgrounds.

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Biology

An Improved Method of RNA Isolation from Loblolly Pine (P. taeda L.) and Other Conifer Species
W. Walter Lorenz 1, Yuan-Sheng Yu 1, Jeffrey F. D. Dean 1
1Warnell School of Forestry and Natural Resources, University of Georgia (UGA)

Many plant tissues, including phloem and xylem from loblolly pine (Pinus taeda L.), contain high levels of phenolics and polysaccharides that interfere with RNA purification. This presentation discusses techniques for the harvest of field-grown tissues and isolation of RNA of sufficient quality for microarrays and other genomic analyses.

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Biology

Bromodeoxyuridine (BrdU) Labeling and Subsequent Fluorescence Activated Cell Sorting for Culture-independent Identification of Dissolved Organic Carbon-degrading Bacterioplankton
Steven Robbins *1, Jisha Jacob *1, Xinxin Lu 1, Mary Ann Moran 2, Xiaozhen Mou 1
1Biological Sciences, Kent State University, 2Marine Sciences, University of Georgia (UGA)

Environmental bacterioplankton are incubated with a model dissolved organic carbon (DOC) compound and a DNA labeling reagent, bromodeoxyuridine (BrdU). Afterward, DOC-degrading cells are separated from the bulk community based on their elevated BrdU incorporation using fluorescence activated cell sorting (FACS). These cells are then identified by subsequent molecular analyses.

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