• Volume 10, 2020
  • Volume 9, 2019
  • Volume 8, 2018
  • Volume 7, 2017
  • Volume 6, 2016
  • Volume 5, 2015
  • Volume 4, 2014
  • Volume 3, 2013
  • Volume 2, 2012
  • Volume 1, 2011
Image Credit: BioProtoc.3792

Cancer Biology

Real-time Three-dimensional Tracking of Endocytic Vesicles Authors:  Stephanie Duhamel and Kossay Zaoui, date: 10/20/2020, view: 525, Q&A: 0
Endocytic trafficking and recycling are fundamental cellular processes that control essential functions such as signaling protein complexes transport and membrane identity. The small GTPase Rabs are indispensable component of the endosomal recycling machinery. The Rabs bind to effectors to mediate their functions, such as protein sorting and ...
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Quantification of Salivary Charged Metabolites Using Capillary Electrophoresis Time-of-flight-mass Spectrometry Authors:  Masahiro Sugimoto, Sana Ota, Miku Kaneko, Ayame Enomoto and Tomoyoshi Soga, date: 10/20/2020, view: 398, Q&A: 0
Salivary metabolomics have provided the potentials to detect both oral and systemic diseases. Capillary electrophoresis time-of-flight-mass spectrometry (CE-TOFMS) enables the identification and quantification of various charged metabolites. This method has been employed to biomarker discoveries using human saliva samples, especially for various ...
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Cell Biology

Live-cell Imaging and Quantitative Analysis of Meiotic Divisions in Caenorhabditis elegans Males Authors:  Gunar Fabig, Falko Löffler, Christian Götze and Thomas Müller-Reichert, date: 10/20/2020, view: 364, Q&A: 0
Live-imaging of meiotic cell division has been performed in extracted spermatocytes of a number of species using phase-contrast microscopy. For the nematode Caenorhabditis elegans, removal of spermatocytes from gonads has damaging effects, as most of the extracted spermatocytes show a high variability in the timing of meiotic divisions or ...
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Microbiology

A Protocol for Simple, Rapid, and Direct Detection of SARS-CoV-2 from clinical samples, using Reverse Transcribed Loop-Mediated Isothermal Amplification (RT-LAMP) Authors:  Rawi Naddaf, Nadav Ben-Assa, Tal Gefen, Tal Capucha, Haitham Hajjo, Noa Mandelbaum, Lilach Elbaum, Shai Kaplan, Assaf Rotem, Michal Chowers, Moran Szwarcwort-Cohen, Mical Paul and Naama Geva-Zatorsky, date: 10/20/2020, view: 492, Q&A: 0
SARS-CoV-2 has quickly spread all around the globe causing illness and wide damages. Most countries were unprepared for such a rapid spread and crisis. This led to various strategies for effective control of the new pandemic. A key aspect in all countries was to effectively test the population for the virus. Most countries chose a lockdown ...
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Estimation of the Minimum Number of Replication Origins Per Chromosome in any Organism Author:  Marcelo S. da Silva, date: 10/20/2020, view: 487, Q&A: 0
Eukaryote nuclear genomes predominantly replicate through multiple replication origins. The number of replication origins activated per chromosome during the S-phase duration may vary according to many factors, but the predominant one is replication stress. Several studies have applied different approaches to estimate the number and map the ...
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Measurement of Ascorbic Acid and Glutathione Content in Cyanobacterium Synechocystis sp. PCC 6803 Authors:  Anabella Aguillera, Charlotte Steelheart, Matías Alegre, Federico Berdun, Graciela Salerno, Carlos Bartoli, Gabriela Pagnussat and María Victoria Martin, date: 10/20/2020, view: 356, Q&A: 0
Ascorbic acid (AsA) and gluthathione (GSH) are two key components of the antioxidant machinery of eukaryotic and prokaryotic cells. The cyanobacterium Synechocystis sp. PCC 6803 presents both compounds in different concentrations (AsA, 20-100 μM and GSH, 2-5 mM). Therefore, it is important to have precise and sensitive methods to ...
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Molecular Biology

Attachment of a 32P-phosphate to the 3′ Terminus of a DNA Oligonucleotide Authors:  Joshua C. Cofsky and Jennifer A. Doudna, date: 10/20/2020, view: 356, Q&A: 0
Biochemical investigations into DNA-binding and DNA-cutting proteins often benefit from the specific attachment of a radioactive label to one of the two DNA termini. In many cases, it is essential to perform two versions of the same experiment: one with the 5′ DNA end labeled and one with the 3′ DNA end labeled. While homogeneous 5′-radiolabeling ...
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Analyzing (Re)Capping of mRNA Using Transcript Specific 5' End Sequencing Authors:  Daniel del Valle Morales and Daniel R. Schoenberg, date: 10/20/2020, view: 354, Q&A: 0
The 5′ cap is a ubiquitous feature of eukaryotic mRNAs. It is added in the nucleus onto newly synthesized pre-mRNA, and in the cytoplasm onto mRNAs after decapping or endonuclease cleavage. Cytoplasmic recapping can occur after loss of the cap at the native 5′ end, or downstream within the body of the mRNA. The identification and location of ...
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Understanding Docking Complexes of Macromolecules Using HADDOCK: The Synergy between Experimental Data and Computations Authors:  Andrea Saponaro, Vincenzo Maione, Alexandre M. J. J. Bonvin and Francesca Cantini, date: 10/20/2020, view: 389, Q&A: 0
This protocol illustrates the modelling of a protein-peptide complex using the synergic combination of in silico analysis and experimental results. To this end, we use the integrative modelling software HADDOCK, which possesses the powerful ability to incorporate experimental data, such as NMR Chemical Shift Perturbations and biochemical ...
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Neuroscience

Staining and Quantitative Analysis of Myelinating Oligodendrocytes in the Mouse Grey Matter Authors:  Matthew Swire and Charles ffrench-Constant, date: 10/20/2020, view: 366, Q&A: 0
Oligodendrocytes generate distinct patterns of myelination throughout the CNS. Variations in myelination along axons may enable neurons to fine-tune conduction velocities and alter signal synchronisation. Here we outline a staining protocol permitting the assessment of the number and length of myelin sheaths formed by oligodendrocyte in the mouse ...
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Plant Science

Acidified Blue Ink-staining Procedure for the Observation of Fungal Structures Inside Roots of Two Disparate Plant Lineages Authors:  Jill Kowal, Elena Arrigoni and Sophie Lane, date: 10/20/2020, view: 319, Q&A: 0
Identifying microscopic mycorrhizal fungal structures in roots, i.e., hyphae, vesicles and arbuscules, requires root staining procedures that are often time consuming and involves chemicals known to present health risks from exposure. By modifying established protocols, our root staining method stains roots using a safe ink- and ...
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Quantification of Methylglyoxal Levels in Cowpea Leaves in Response to Cowpea Aphid Infestation Authors:  Jacob R. MacWilliams, Monika Ostaszewska-Bugajska, Klaudia Borysiuk, Bożena Szal and Isgouhi Kaloshian, date: 10/20/2020, view: 247, Q&A: 0
Aphids are a serious pest of crops across the world. Aphids feed by inserting their flexible hypodermal needlelike mouthparts, or stylets, into their host plant tissues. They navigate their way to the phloem where they feed on its sap causing little mechanical damage to the plant. Additionally, while feeding, aphids secrete proteinaceous effectors ...
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Dual sgRNA-based Targeted Deletion of Large Genomic Regions and Isolation of Heritable Cas9-free Mutants in Arabidopsis Authors:  Yu Jin and Sebastian Marquardt, date: 10/20/2020, view: 523, Q&A: 0
CRISPR/Cas9 system directed by a gene-specific single guide RNA (sgRNA) is an effective tool for genome editing such as deletions of few bases in coding genes. However, targeted deletion of larger regions generate loss-of-function alleles that offer a straightforward starting point for functional dissections of genomic loci. We present an ...
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Low-cost and High-throughput RNA-seq Library Preparation for Illumina Sequencing from Plant Tissue Authors:  Marta Bjornson, Kaisa Kajala, Cyril Zipfel and Pingtao Ding, date: 10/20/2020, view: 686, Q&A: 0
Transcriptome analysis can provide clues to biological processes affected in different genetic backgrounds or/and under various conditions. The price of RNA sequencing (RNA-seq) has decreased enough so that medium- to large-scale transcriptome analyses in a range of conditions are feasible. However, the price and variety of options for library ...
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Stem Cell

Derivation of Induced Pluripotent Stem Cells from Human Fibroblasts Using a Non-integrative System in Feeder-free Conditions Authors:  Alvaro A. Beltran, Sarahi G. Molina and Adriana S. Beltran, date: 10/20/2020, view: 324, Q&A: 0
Induced pluripotent stem cells (iPSCs) are genetically reprogrammed somatic cells that exhibit features identical to those of embryonic stem cells (ESCs). Multiple approaches are available to derive iPSCs, among which the Sendai virus is the most effective at reprogramming different cell types. Here we describe a rapid, efficient, safe, and ...
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