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Latest Articles

Microscopic Investigation of Protein Function in C. elegans Using Fluorescent Imaging
Cliff J. Luke, Linda P. O'Reilly
Caenorhabditis elegans is a powerful model organism for studying human biology and disease due to its surprisingly high genetic homology to Homo sapiens . Its genetic amenability, small size, short generation time, and transparent body make it an ideal organism for multiple scientific disciplines.
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In Vivo Immuno‐Spin Trapping: Imaging the Footprints of Oxidative Stress
Nicholas K.H. Khoo, Nadiezhda Cantu‐Medellin, Claudette St. Croix, Eric E. Kelley
A plethora of disease processes are associated with elevated reactive species formation and allied reactions with biomolecules that alter cell signaling, induce overt damage, and promote dysfunction of tissues. Unfortunately, effective detection of reactive species in tissues is wrought with issues
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Transcriptome‐Wide Identification of Pseudouridine Modifications Using Pseudo‐seq
Thomas M. Carlile, Maria F. Rojas‐Duran, Wendy V. Gilbert
A diverse array of post‐transcriptional modifications is found in RNA molecules from all domains of life. While the locations of RNA modifications are well characterized in abundant noncoding RNAs, modified sites in less abundant mRNAs are just beginning to be discovered. Recent work has revealed
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Overview of Target Enrichment Strategies
Iwanka Kozarewa, Javier Armisen, Andrew F. Gardner, Barton E. Slatko, C.L. Hendrickson
Target enrichment is commonly used in next generation sequencing (NGS) workflows to eliminate genomic DNA regions that are not of interest for a particular experiment. By only targeting specific regions such as exons, one can obtain greater depth of DNA sequencing coverage for regions of interest or
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Laser Microdissection
Andra R. Frost, Isam‐Eldin Eltoum, Gene P. Siegal, Michael R. Emmert‐Buck, Michael A. Tangrea
Laser microdissection (LM) offers a relatively rapid and precise method of isolating and removing specified cells from complex tissues for subsequent analysis of their RNA, DNA, protein or metabolite content, thereby allowing assessment of the role of different cell types in the normal physiological
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Laser Microdissection–Mediated Isolation and In Vitro Transcriptional Amplification of Plant RNA
Divya Chandran, Michael J. Scanlon, Kazuhiro Ohtsu, Marja C.P. Timmermans, Patrick S. Schnable, Mary C. Wildermuth
Laser microdissection of cells allows for isolation of specific cells of interest for downstream analyses including transcriptional profiling. Plant cells present unique challenges for laser microdissection due to their cellulosic cell walls and large vacuoles. Here we present protocols for plant
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RNA Interference and MicroRNA‐Mediated Silencing
Sylvia E.J. Fischer
RNA interference (RNAi) and microRNA‐mediated silencing cause sequence‐specific silencing of target genes. This overview will give a brief description of how RNAi and microRNAs were discovered, how small RNAs silence their targets, and what the functions of small RNAs are. Since the discovery of
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Genome Editing in Human Cells Using CRISPR/Cas Nucleases
Nicolas Wyvekens, Shengdar Q. Tsai, J. Keith Joung
The clustered regularly interspaced short palindromic repeat (CRISPR)/CRISPR‐associated (Cas) system has been broadly adopted for highly efficient genome editing in a variety of model organisms and human cell types. Unlike previous genome editing technologies such as zinc finger nucleases (ZFNs) and
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Whole Mouse Brain Image Reconstruction from Serial Coronal Sections Using FIJI (ImageJ)
Ronald Paletzki, Charles R. Gerfen
Whole‐brain reconstruction of the mouse enables comprehensive analysis of the distribution of neurochemical markers, the distribution of anterogradely labeled axonal projections or retrogradely labeled neurons projecting to a specific brain site, or the distribution of neurons displaying
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Laser‐Mediated Microlesions in Mouse Neocortex to Investigate Neuronal Degeneration and Regeneration
Johanna Jackson, Alison J. Canty, Lieven Huang, Vincenzo De Paola
In vivo two‐photon (2P) imaging enables neural circuitry to be repeatedly visualized in both normal conditions and following trauma. This protocol describes how laser‐mediated neuronal microlesions can be created in the cerebral cortex using an ultrafast laser without causing a significant
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Use of Adeno‐Associated and Herpes Simplex Viral Vectors for In Vivo Neuronal Expression in Mice
Rachel D. Penrod, Audrey M. Wells, William A. Carlezon, Christopher W. Cowan
Adeno‐associated viruses and the herpes simplex virus are the two most widely used vectors for the in vivo expression of exogenous genes. Advances in the development of these vectors have enabled remarkable temporal and spatial control of gene expression. This unit provides methods for storing,
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