Molecular Biology

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    Protocols in Current Issue
    Quantification of HIV-2 DNA in Whole Blood
    Authors:  Zsófia Szojka, Sara Karlson, Marianne Jansson and Patrik Medstrand, date: 10/20/2019, view: 270, 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 ...
    ATAC-seq on Sorted Adult Mouse Neurons
    Authors:  Erin A. Clark, Yasuyuki Shima and Sacha Nelson, date: 10/05/2019, view: 404, Q&A: 0
    [Abstract] Transcription regulation is a key aspect of cellular identity established during development and maintained into adulthood. Molecular and biochemical assays that probe the genome are critical tools in exploring mechanisms of transcription regulation and cell type identity. The mammalian brain is composed of a huge diversity of cell types with ...
    In situ Hybridization of Plant-parasitic Nematode Globodera pallida Juveniles to Detect Gene Expression
    [Abstract] In this study, we describe a standard whole mount in situ hybridization method which is used to determine the spatial-temporal expression pattern of genes from Globodera spp. Unlike more invasive radioactive labeling approaches, this technique is based on a safe, highly specific enzyme-linked immunoassay where a Digoxigenin ...
    Using indCAPS to Detect CRISPR/Cas9 Induced Mutations
    Authors:  Charles Hodgens, Zachary L. Nimchuk and Joseph J. Kieber, date: 09/20/2019, view: 250, Q&A: 0
    [Abstract] Cleaved amplified polymorphic sequences (CAPS) assays are useful tools for detecting small mutations such as single nucleotide polymorphisms (SNPs) or insertion/deletions (indels) present in an amplified DNA fragment. A mutation that disrupts or creates a restriction site will prevent cleavage by a restriction enzyme, allowing discrimination of ...
    A Refined Protocol for Identifying Citrulline-specific Monoclonal Antibodies from Single Human B Cells from Rheumatoid Arthritis Patient Material
    [Abstract] We describe here a detailed, refined protocol for the generation of citrulline-specific monoclonal antibodies from single human B cells from rheumatoid arthritis (RA) patients. This protocol provides a detailed guide of the procedure starting from single B cells of your choice and followed by amplification of the variable region of immunoglobulin ...
    PEG-mediated, Stable, Nuclear and Chloroplast Transformation of Cyanidioschizon merolae
    Authors:  Maksymilian Zienkiewicz, Tomasz Krupnik, Anna Drożak and Kinga Kania, date: 09/05/2019, view: 325, Q&A: 0
    [Abstract] The ability to achieve nuclear or chloroplast transformation in plants has been a long standing goal, especially in microalgae research. Over past years there has been only little success, but transient and stable nuclear transformation has been achieved in multiple species. Our newly developed method allows for relatively simple transformation of ...
    Development of an Efficient Transformation System for Halotolerant Yeast Debaryomyces hansenii CBS767
    Authors:  Anu P. Minhas and Dipanwita Biswas, date: 09/05/2019, view: 292, Q&A: 0
    [Abstract] Debaryomyces hansenii is one of the most osmotolerant and halotolerant yeasts. Further, its association with traditional cheese and meat products imparting special flavors to these products project this yeast with enormous biotechnological potential in the agrofood sector. However, lack of an efficient transformation system in D. hansenii ...
    A Novel Dual Lentiviral CRISPR-based Transcriptional Activation System for Gene Expression Regulation in Neurons
    Authors:  Katherine E. Savell, Faraz A. Sultan and Jeremy J. Day, date: 09/05/2019, view: 834, Q&A: 0
    [Abstract] Robust and efficient gene expression control enables the study of a gene’s function in the central nervous system. Advances in CRISPR-based technology provide new avenues not only for gene editing, but for complex transcriptional control. Here, we describe a protocol to generate high-titer lentiviruses with neuron-optimized CRISPR-activation ...
    Construction of Viral Vectors for Cell Type-specific CRISPR Gene Editing in the Adult Mouse Brain
    Authors:  Hiroshi Yamaguchi and Luis de Lecea, date: 08/20/2019, view: 710, Q&A: 0
    [Abstract] Recently developed gene editing technologies based on engineered CRISPR/Cas9 systems enables researchers to disrupt genes in a cell type-specific manner in the adult mouse brain. Using these technologies, we recently showed that the dopamine beta-hydroxylase gene in Locus Coeruleus (LC) norepinephrine neurons plays a vital role in the maintenance ...



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