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Colorado State University

3 ARTICLES PUBLISHED IN JoVE

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Biology

In vivo Electroporation of Morpholinos into the Regenerating Adult Zebrafish Tail Fin
David R. Hyde 1, Alan R. Godwin 2, Ryan Thummel 3
1Department of Biological Sciences, Center for Zebrafish Research, University of Notre Dame , 2Department of Microbiology, Immunology, and Pathology, Colorado State University , 3Departments of Anatomy and Cell Biology and Ophthalmology, Wayne State University School of Medicine

We describe a method to conditionally knockdown the expression of a target protein during adult zebrafish fin regeneration. This technique involves micro-injecting and electroporating antisense oligonucleotide morpholinos into fin tissue, which allows testing the protein’s role in various stages of fin regeneration, including wound healing, blastema formation, and regenerative outgrowth.

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JoVE Core

Assembly of Nucleosomal Arrays from Recombinant Core Histones and Nucleosome Positioning DNA
Ryan A. Rogge 1, Anna A. Kalashnikova 1, Uma M. Muthurajan 1, Mary E. Porter-Goff 1, Karolin Luger 1, Jeffrey C. Hansen 1
1Biochemistry and Molecular Biology, Colorado State University

A method is presented for the reconstitution of model nucleosomal arrays from recombinant core histones and tandemly repeated nucleosome positioning DNA. We also describe how sedimentation velocity experiments in the analytical ultracentrifuge, and atomic force microscopy (AFM) are used to monitor the extent of nucleosomal array saturation after reconstitution.

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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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