March 2015: This Month in JoVE - Solving Crime with Science, Applying Technology to Understand Trees, and Studying Protein Synthesis on a Chip

Protocol

Dried Blood Spots — Preparing and Processing for Use in Immunoassays and in Molecular Techniques

Nico Grüner, Oumaima Stambouli, R. Stefan Ross

Institute of Virology, National Reference Centre for Hepatitis C, Essen University Hospital, University of Duisburg-Essen

The preparing and processing of dried blood spots (DBS) for their final analysis are still poorly standardized for most diagnostic applications. To overcome this shortcoming, a comprehensive step-by-step protocol is suggested and subsequently evaluated with regard to its effectiveness for detecting markers of viral infections.

A Technical Perspective in Modern Tree-ring Research — How to Overcome Dendroecological and Wood Anatomical Challenges

Holger Gärtner, Paolo Cherubini, Patrick Fonti, Georg von Arx, Loïc Schneider, Daniel Nievergelt, Anne Verstege, Alexander Bast, Fritz H. Schweingruber, Ulf Büntgen

Landscape Dynamics / Dendroecology, Swiss Federal Research Institute WSL

Here we present a protocol outlining how to sample wooden specimens for the overall assessment of their growth structures. Macro- and microscopic preparation and visualization techniques necessary to generate well-replicated and highly resolved wood anatomical and dendroecological dataset, are described are described.

Enhanced Genetic Analysis of Single Human Bioparticles Recovered by Simplified Micromanipulation from Forensic ‘Touch DNA’ Evidence

Katherine Farash^1,2, Erin K. Hanson³, Jack Ballantyne^1,2,3

¹Forensic Science Graduate Program, Biochemistry Track, University of Central Florida, ²Department of Chemistry, University of Central Florida, ³National Center for Forensic Science, University of Central Florida

Here we describe an optimized and efficient removal strategy for the collection of bio-particles present in ‘touch DNA’ samples, together with an enhanced amplification protocol involving a one-step 5 µl micro-volume lysis/STR amplification, to permit the recovery of short tandem repeat (STR) profiles of the bio-particle donor(s).

Sealable Femtoliter Chamber Arrays for Cell-free Biology

Sarah Elizabeth Norred^1,2, Patrick M. Caveney^1,2, Scott T. Retterer^1,2, Jonathan B. Boreyko^1,2, Jason D. Fowlkes^2,3, Charles Patrick Collier², Michael L. Simpson^1,2,3

¹Bredesen Center, University of Tennessee, Knoxville, ²Center for Nanophase Materials Sciences, Oak Ridge National Laboratory, ³Department of Materials Science and Engineering, University of Tennessee, Knoxville

A microfabricated device with sealable femtoliter-volume reaction chambers is described. This report includes a protocol for sealing cell-free protein synthesis reactants inside these chambers for the purpose of understanding the role of crowding and confinement in gene expression.