Molecular Biology

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    Protocols in Current Issue
    SarkoSpin: A Technique for Biochemical Isolation and Characterization of Pathological TDP-43 Aggregates
    Authors:  Pérez-Berlanga Manuela, Laferrière Florent and Polymenidou Magdalini, date: 11/20/2019, view: 198, Q&A: 0
    TDP-43 is the main aggregating protein in neurodegenerative diseases such as amyotrophic lateral sclerosis (ALS) and frontotemporal lobar degeneration (FTLD). Aggregated TDP-43 is resistant to diverse detergent solubilization, yet physiological ...
    ELISA Based Protein Ubiquitylation Measurement
    Authors:  Yuka Kamada, Ryosuke Fukuda and Tsukasa Okiyoneda, date: 11/20/2019, view: 22, Q&A: 0
    Ubiquitylation is a common post-translational modification of cellular proteins that results in proteasomal and lysosomal degradations. Ubiquitylation is generally measured by methods such as immunoblotting using anti-ubiquitin antibodies after ...
    Determination of Chromatin Accessibility in Drosophila Midgut Enterocytes by in situ 5mC Labeling
    Authors:  Eliya Bitman-Lotan, Hector Rincon-Arano, Gal Raz and Amir Orian, date: 11/20/2019, view: 20, Q&A: 0
    Regulation of gene expression involves dynamic changes in chromatin organization, where in many cases open chromatin structure correlates with gene activation. Several methods enable monitoring changes in chromatin accessibility, including ATAC-seq, ...
    Labeling and Isolation of Fluorouracil Tagged RNA by Cytosine Deaminase Expression
    Authors:  Harihar Basnet and Joan Massague, date: 11/20/2019, view: 28, Q&A: 0
    Tissues are comprised of different cell types whose interactions elicit distinct gene expression patterns that regulate tissue formation, regeneration, homeostasis and repair. Analysis of these gene expression patterns require methods that can ...
    SarkoSpin: A Technique for Biochemical Isolation and Characterization of Pathological TDP-43 Aggregates
    Authors:  Pérez-Berlanga Manuela, Laferrière Florent and Polymenidou Magdalini, date: 11/20/2019, view: 198, Q&A: 0
    [Abstract] TDP-43 is the main aggregating protein in neurodegenerative diseases such as amyotrophic lateral sclerosis (ALS) and frontotemporal lobar degeneration (FTLD). Aggregated TDP-43 is resistant to diverse detergent solubilization, yet physiological TDP-43 and other abundant proteins commonly co-purify with pathological TDP-43. This mixed isolation has ...
    ELISA Based Protein Ubiquitylation Measurement
    Authors:  Yuka Kamada, Ryosuke Fukuda and Tsukasa Okiyoneda, date: 11/20/2019, view: 22, Q&A: 0
    [Abstract] Ubiquitylation is a common post-translational modification of cellular proteins that results in proteasomal and lysosomal degradations. Ubiquitylation is generally measured by methods such as immunoblotting using anti-ubiquitin antibodies after isolating the protein-of-interest by denaturing immunoprecipitation. The following protocol can be used ...
    Determination of Chromatin Accessibility in Drosophila Midgut Enterocytes by in situ 5mC Labeling
    Authors:  Eliya Bitman-Lotan, Hector Rincon-Arano, Gal Raz and Amir Orian, date: 11/20/2019, view: 20, Q&A: 0
    [Abstract] Regulation of gene expression involves dynamic changes in chromatin organization, where in many cases open chromatin structure correlates with gene activation. Several methods enable monitoring changes in chromatin accessibility, including ATAC-seq, FAIRE-seq, MNase-seq and DNAse-seq methods, which involve Next-generation-sequencing (NGS). ...
    Labeling and Isolation of Fluorouracil Tagged RNA by Cytosine Deaminase Expression
    Authors:  Harihar Basnet and Joan Massague, date: 11/20/2019, view: 28, Q&A: 0
    [Abstract] Tissues are comprised of different cell types whose interactions elicit distinct gene expression patterns that regulate tissue formation, regeneration, homeostasis and repair. Analysis of these gene expression patterns require methods that can capture as closely as possible the transcriptomes of cells of interest in their tissue microenvironment. ...
    Using Imaging Flow Cytometry to Characterize Extracellular Vesicles Isolated from Cell Culture Media, Plasma or Urine
    Authors:  John R. Woollard, Amrutesh Puranik, Kyra L. Jordan and Lilach O. Lerman, date: 11/05/2019, view: 162, Q&A: 0
    [Abstract] The ability to non-invasively detect specific damage to the kidney has been limited. Identification of extracellular vesicles released by cells, especially when under duress, might allow for monitoring and identification of specific cell types within the kidney that are stressed. We have adapted a previously published traditional flow cytometry ...
    A Mismatch-tolerant RT-LAMP Method for Molecular Diagnosis of Highly Variable Viruses
    Authors:  Yingxue Li, Yi Zhou, Yingying Ma, Rong Xu, Xia Jin and Chiyu Zhang, date: 11/05/2019, view: 162, Q&A: 0
    [Abstract] Loop-mediated isothermal amplification (LAMP) has been widely used in the detection of pathogens. However, there are usually numerous variants in one viral pathogen and primers employed in LAMP can hardly match all these variants. The mismatches between the primers and the viral genomes, especially those at the 3′-end of the primers, hinder LAMP ...
    Platelet Isolation and Activation Assays
    Authors:  Laura C. Burzynski, Nicholas Pugh and Murray C.H. Clarke, date: 10/20/2019, view: 524, Q&A: 0
    [Abstract] Platelets regulate hemostasis and are the key determinants of pathogenic thrombosis following atherosclerotic plaque rupture. Platelets circulate in an inactive state, but become activated in response to damage to the endothelium, which exposes thrombogenic material such as collagen to the blood flow. Activation results in a number of responses, ...
    Quantification of HIV-2 DNA in Whole Blood
    Authors:  Zsófia Szojka, Sara Karlson, Marianne Jansson and Patrik Medstrand, date: 10/20/2019, view: 298, Q&A: 0
    [Abstract] Time to AIDS infection is longer with HIV-2, compared to HIV-1, but without antiretroviral therapy both infections will cause AIDS-related mortality. In HIV-2 infection, monitoring of antiretroviral treatment (ART) efficacy is challenging since a large proportion of HIV-2-infected individuals displays low or undetectable plasma RNA levels. Hence, ...
    Simple Synthesis of Functionalized Paramagnetic Beads for Nucleic Acid Purification and Manipulation
    [Abstract] The purification of nucleic acids is one of the most common procedures employed in modern molecular biology laboratories. Typically, commercial column-based protocols are utilized to isolate DNA or RNA from various sources. However, these methods not only require specialized equipment, but are also extremely expensive for high-throughput ...
    Isolation, Purification, and Characterization of Ginger-derived Nanoparticles (GDNPs) from Ginger, Rhizome of Zingiber officinale
    Authors:  Junsik Sung, Chunhua Yang, Emilie Viennois, Mingzhen Zhang and Didier Merlin, date: 10/05/2019, view: 501, Q&A: 1
    [Abstract] Factors implicated in the pathophysiology of intestinal inflammation include defects in intestinal epithelial barrier function, abnormal immune responses, and activities of the gut microbiota. Current agents used to treat human Inflammatory Bowels Disease (IBD), chronic inflammation of digestive tract, have serious side effects. In addition, most ...



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