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University of Wisconsin - Madison

5 ARTICLES PUBLISHED IN JoVE

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Biology

GC-based Detection of Aldononitrile Acetate Derivatized Glucosamine and Muramic Acid for Microbial Residue Determination in Soil
Chao Liang 1,2, Harry W. Read 2, Teri C. Balser 2,3
1DOE-Great Lakes Bioenergy Research Center, University of Wisconsin, Madison, 2Department of Soil Science, University of Wisconsin, Madison, 3Department of Soil and Water Science, University of Florida

We describe a method protocol for the GC-based analysis of the aldonitrile acetate derivatives of glucosamine and muramic acid extracted from soil. For elucidation of the chemical mechanism, we also present a strategy to confirm the structure of the derivative and the ion fragments formed upon electron ionization.

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Biology

Monitoring Plasmid Replication in Live Mammalian Cells over Multiple Generations by Fluorescence Microscopy
Kathryn Norby 1, Ya-Fang Chiu 1, Bill Sugden 1
1Department of Oncology, University of Wisconsin - Madison

A method of observing individual DNA molecules in live cells is described. The technique is based on the binding of a fluorescently tagged lac repressor protein to binding sites engineered into the DNA of interest. This method can be adapted to follow many recombinant DNAs in live cells over time.

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Chemistry

Rapid High Throughput Amylose Determination in Freeze Dried Potato Tuber Samples
Diego Fajardo 1, Sastry S. Jayanty 2, Shelley H. Jansky 1
1USDA-ARS and Department of Horticulture, University of Wisconsin - Madison, 2Department of Horticulture and Landscape Architecture, Colorado State University

This protocol describes a high through put colorimetric method that relies on the formation of a complex between iodine and chains of glucose molecules in starch. Iodine forms complexes with both amylose and long chains within amylopectin. After the addition of iodine to a starch sample, the maximum absorption of amylose and amylopectin occurs at 620 and 550 nm, respectively. The amylose/amylopectin ratio can be estimated from the ratio of the 620 and 550 nm absorbance values and comparing them to a standard curve in which specific known concentrations are plotted against absorption values. This high throughput, inexpensive method is reliable and reproducible, allowing the evaluation of large populations of potato clones. 

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JoVE Core

Measurement of Greenhouse Gas Flux from Agricultural Soils Using Static Chambers
Sarah M. Collier 1, Matthew D. Ruark 2, Lawrence G. Oates 3,4, William E. Jokela 5, Curtis J. Dell 6
1Office of Sustainability, University of Wisconsin-Madison, 2Department of Soil Science, University of Wisconsin-Madison, 3Department of Agronomy, University of Wisconsin-Madison, 4Great Lakes Bioenergy Research Center, University of Wisconsin-Madison, 5USDA-ARS Dairy Forage Research Center, 6USDA-ARS Pasture Systems Watershed Management Research Unit

This article showcases the static chamber-based method for measurement of greenhouse gas flux from soil systems. With relatively modest infrastructure investments, measurements may be obtained from multiple treatments/locations and over timeframes ranging from hours to years.

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Environment

A Lipid Extraction and Analysis Method for Characterizing Soil Microbes in Experiments with Many Samples
Lawrence G. Oates 1, Harry W. Read 2, Jessica L. M. Gutknecht 3, David S. Duncan 1, Teri B. Balser 4, Randall D. Jackson 1
1Department of Agronomy and Great Lakes Bioenergy Research Center, University of Wisconsin - Madison, 2Department of Soil Science, University of Wisconsin - Madison, 3Department of Soil, Water, and Climate, University of Minnesota, 4Faculty of Science and Engineering, Curtin University

The article describes a method that increases throughput while balancing effort and accuracy for extraction of lipids from the cell membranes of microorganisms for use in characterizing both total lipids and the relative abundance of indicator lipids to determine soil microbial community structure in studies with many samples.

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