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JoVE Biology
 JoVE Biology

A Method for Selecting Structure-switching Aptamers Applied to a Colorimetric Gold Nanoparticle Assay

1711th Human Performance Wing, Human Effectiveness Directorate, Air Force Research Laboratory, Wright-Patterson Air Force Base, 2The Henry M. Jackson Foundation, 3UES, Inc.


JoVE 52545

A protocol is provided to select structure-switching aptamers for small molecule targets based on a tunable stringency magnetic bead selection method. Aptamers selected with structure-switching properties are beneficial for assays that require a conformational change to signal the presence of a target, such as the described gold nanoparticle assay.

 JoVE Biology

A Multi-detection Assay for Malaria Transmitting Mosquitoes

1Department of Pathology, Microbiology and Immunology, School of Veterinary Medicine, University of California - Davis, 2Veterinary Genetics Laboratory, School of Veterinary Medicine, University of California, Davis


JoVE 52385

Malaria transmitting mosquitoes have a number of epidemiologically important characteristics that can only be detected using molecular techniques. Utilizing a MALDI-TOF based SNP genotyping platform, we developed an assay for simultaneously detecting multiple key traits (species, insecticide resistance, parasite infection and host choice) of malaria vectors.

 JoVE Biology

Targeted DNA Methylation Analysis by Next-generation Sequencing

1Department of Physiology, University of Oklahoma College of Medicine, 2Department of Geriatric Medicine, University of Oklahoma College of Medicine


JoVE 52488

Bisulfite amplicon sequencing (BSAS) is a method for quantifying cytosine methylation in targeted genomic regions of interest. This method uses bisulfite conversion paired with PCR amplification of target regions prior to next-generation sequencing to produce absolute quantitation of DNA methylation at a base-specific level.

 JoVE Biology

Enhanced Reduced Representation Bisulfite Sequencing for Assessment of DNA Methylation at Base Pair Resolution

1Department of Medicine, Weill Cornell Medical College, 2Institute for Computational Biomedicine, Weill Cornell Medical College, 3Department of Physiology and Biophysics, Weill Cornell Medical College, 4Department of Pathology, University of Michigan


JoVE 52246

Enhanced Reduced Representation Bisulfite Sequencing is a method for the preparation of sequencing libraries for DNA methylation analysis based on restriction enzyme digestion combined with cytosine bisulfite conversion. This protocol requires 50 ng of starting material and yields base pair resolution data at GC-rich genomic regions.

 JoVE Biology

Array Comparative Genomic Hybridization (Array CGH) for Detection of Genomic Copy Number Variants

1Cytogenetics Department, Guy's & St Thomas' NHS Foundation Trust, 2Cytogenetics Department, Viapath Analytics


JoVE 51718

Array CGH for the detection of genomic copy number variants has replaced G-banded karyotype analysis. This paper describes the technology and its application in a diagnostic service laboratory.

 JoVE Biology

Evaluation of Zebrafish Kidney Function Using a Fluorescent Clearance Assay

1Genetics and Genomic Medicine, Institute of Child Health, University College London, 2Molecular Cell Science Research Centre, St. George's University of London


JoVE 52540

The zebrafish is a popular tool to model chronic kidney disease (CKD). However, their small size makes it impossible to evaluate renal function using traditional methods. We describe a fluorescent dye kidney clearance assay1 that allows quantitative analysis of zebrafish kidney function in CKD.

 JoVE Biology

Growth-based Determination and Biochemical Confirmation of Genetic Requirements for Protein Degradation in Saccharomyces cerevisiae

1Department of Biology, Ball State University, 2Division of Nephrology, Cincinnati Children's Hospital


JoVE 52428

This article describes a yeast growth-based assay for the determination of genetic requirements for protein degradation. It also demonstrates a method for rapid extraction of yeast proteins, suitable for western blotting to biochemically confirm degradation requirements. These techniques can be adapted to monitor degradation of a variety of proteins.

 JoVE Biology

Strategies for Tracking Anastasis, A Cell Survival Phenomenon that Reverses Apoptosis

1W. Harry Feinstone Department of Molecular Microbiology and Immunology, Johns Hopkins University Bloomberg School of Public Health, 2School of Life Sciences, Chinese University of Hong Kong, 3Center for Cell Dynamics, Department of Biological Chemistry, Johns Hopkins University School of Medicine


JoVE 51964

The term anastasis refers to the phenomenon in which dying cells reverse a cell suicide process at a late stage, repair themselves, and ultimately survive. Here we demonstrate protocols for detecting and tracking cells that undergo anastasis.

 JoVE Biology

Generation of Plasmid Vectors Expressing FLAG-tagged Proteins Under the Regulation of Human Elongation Factor-1α Promoter Using Gibson Assembly

1Department of Cell Biology and Biochemistry, Texas Tech University Health Sciences Center


JoVE 52235

Synthesis of custom plasmids is labor and time consuming. This protocol describes the use of Gibson assembly cloning to reduce the work and duration of custom DNA cloning procedure. The protocol described also produces reliable tagged protein constructs for mammalian expression at similar cost to the traditional cut-and-paste DNA cloning.

 JoVE Biology

Internalization and Observation of Fluorescent Biomolecules in Living Microorganisms via Electroporation

1Clarendon Laboratory, Department of Physics, University of Oxford, 2Wellcome Trust Sanger Institute, Genome Center


JoVE 52208

Studies of biomolecules in vivo are crucial for understanding molecular function in a biological context. Here we describe a novel method allowing the internalization of fluorescent biomolecules, such as DNA or proteins, into living microorganisms. Analysis of in vivo data recorded by fluorescence microscopy is also presented and discussed.

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