Molecular Biology

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    Protocols in Current Issue
    Detection of Ligand-binding to Membrane Proteins by Capacitance Measurements
    Authors:  Verena Burtscher, Matej Hotka and Walter Sandtner, date: 01/05/2019, view: 180, Q&A: 0
    In multi-cellular organisms, cells communicate with each other utilizing chemical messengers. For many of these messenger molecules, the membrane is an insurmountable barrier. Yet, they act by binding to surface proteins often triggering a cascade ...
    In vitro Generation of CRISPR-Cas9 Complexes with Covalently Bound Repair Templates for Genome Editing in Mammalian Cells
    The CRISPR-Cas9 system is a powerful genome-editing tool that promises application for gene editing therapies. The Cas9 nuclease is directed to the DNA by a programmable single guide (sg)RNA, and introduces a site-specific double-stranded break ...
    Gene Mapping by RNA-sequencing: A Direct Way to Characterize Genes and Gene Expression through Targeted Queries of Large Public Databases
    Author:  Peter Rotwein, date: 01/05/2019, view: 215, Q&A: 0
    Recent advances in genomics present new opportunities for enhancing knowledge about gene regulation and function across a wide spectrum of organisms and species. Understanding and evaluating this information at the individual gene level is ...
    Detection of Ligand-binding to Membrane Proteins by Capacitance Measurements
    Authors:  Verena Burtscher, Matej Hotka and Walter Sandtner, date: 01/05/2019, view: 180, Q&A: 0
    [Abstract] In multi-cellular organisms, cells communicate with each other utilizing chemical messengers. For many of these messenger molecules, the membrane is an insurmountable barrier. Yet, they act by binding to surface proteins often triggering a cascade of reactions inside the cell. Accordingly, studying ligand-receptor interactions at the cellular ...
    In vitro Generation of CRISPR-Cas9 Complexes with Covalently Bound Repair Templates for Genome Editing in Mammalian Cells
    [Abstract] The CRISPR-Cas9 system is a powerful genome-editing tool that promises application for gene editing therapies. The Cas9 nuclease is directed to the DNA by a programmable single guide (sg)RNA, and introduces a site-specific double-stranded break (DSB). In mammalian cells, DSBs are either repaired by non-homologous end joining (NHEJ), generating ...
    Gene Mapping by RNA-sequencing: A Direct Way to Characterize Genes and Gene Expression through Targeted Queries of Large Public Databases
    Author:  Peter Rotwein, date: 01/05/2019, view: 215, Q&A: 0
    [Abstract] Recent advances in genomics present new opportunities for enhancing knowledge about gene regulation and function across a wide spectrum of organisms and species. Understanding and evaluating this information at the individual gene level is challenging, and not only requires extracting, collating and interpreting data from public genetic ...
    Production, Titration and Imaging of Zika Virus in Mammalian Cells
    Authors:  Wesley Freppel, Clément Mazeaud and Laurent Chatel-Chaix, date: 12/20/2018, view: 344, Q&A: 0
    [Abstract] Since the outbreak of Zika virus (ZIKV) in Latin America and the US in 2016, this flavivirus has emerged as a major threat for public health. Indeed, it is now clear that ZIKV is vertically transmitted from the infected mother to the fetus and this may lead to severe neurological development defects including (but not restricted to) neonate ...
    Variable Dose Analysis: A Novel High-throughput RNAi Screening Method for Drosophila Cells
    Authors:  Katarzyna Sierzputowska, Chris R. Baxter and Benjamin E. Housden, date: 12/20/2018, view: 334, Q&A: 0
    [Abstract] Genetic screens are a powerful approach to identify previously uncharacterized genes involved in specific biological processes. Several technologies have been developed for high-throughput screens using reagents such as RNAi or CRISPR, and each approach is associated with specific advantages and disadvantages. Variable Dose Analysis (VDA), is an ...
    Visualization and Quantification of Cell-to-cell Movement of Proteins in Nicotiana benthamiana
    Authors:  Mila C Blekemolen, Mariliis Tark-Dame and Frank L.W Takken, date: 12/20/2018, view: 446, Q&A: 0
    [Abstract] Cell-to-cell movement of proteins through plasmodesmata is a widely-established mechanism for intercellular signaling in plants. Current techniques to study intercellular protein translocation rely on single-cell transformation using particle bombardment or transgenic lines expressing photo-inducible fluorophores. The method presented here allows ...
    Analysis of DNA Exchange Using Thymidine Analogs (ADExTA) in Trypanosoma cruzi
    [Abstract] Trypanosoma cruzi is a protozoan parasite belonging to the Trypanosomatidae family. Although the trypanosomatids multiply predominantly by clonal generation, the presence of DNA exchange in some of them has been puzzling researchers over the years, mainly because it may represent a novel form that these organisms use to gain variability. ...
    Cluster Analysis of Endogenous HER2 and HER3 Receptors in SKBR3 Cells
    [Abstract] The Human Epidermal Growth Factor Receptor (HER) family of receptor tyrosine kinases consists of four, single pass, transmembrane receptor homologs (HER1-4) that act to regulate many critical processes in normal and tumor cells. HER2 is overexpressed in many tumors, and the deregulated proliferation of cancerous cells is driven by cooperation with ...
    Quantitative Transformation Efficiency Assay for Bacillus subtilis
    Authors:  Christian L. Loyo and Briana M. Burton, date: 12/05/2018, view: 344, Q&A: 0
    [Abstract] Bacillus subtilis (B. subtilis) is a model Gram-positive organism used to study a variety of physiological states and stress responses, one of which is the development of competence. Competence refers to the physiological state of a cell which allows it to be transformed naturally. Through induction of competence, the efficiency ...
    Wheat Germ Agglutinin (WGA)-SDS-PAGE: A Novel Method for the Detection of O-GlcNAc-modified Proteins by Lectin Affinity Gel Electrophoresis
    Authors:  Ko Fujioka, Yuji Kubota and Mutsuhiro Takekawa, date: 12/05/2018, view: 393, Q&A: 0
    [Abstract] Diverse cytoplasmic and nuclear proteins dynamically change their molecular functions by O-linked β-N-acetylglucosamine (O-GlcNAc) modification on serine and/or threonine residues. Evaluation of the O-GlcNAcylation level of a specific protein, however, needs multiple and time-consuming steps if using conventional methods (e.g., ...



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