Molecular Biology

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    Protocols in Current Issue
    Transcervical Mouse Infections with Chlamydia trachomatis and Determination of Bacterial Burden
    Authors:  Karthika Rajeeve and Rajeeve Sivadasan, date: 02/05/2020, view: 168, Q&A: 0
    [Abstract] Chlamydia trachomatis is an obligate human pathogen. It infects the genital tract of humans ascending into the fallopian tube, exacerbated by chronic pelvic pain, pelvic inflammatory disease, and fallopian tube scaring resulting in infertility and other malignancies. The major hurdle in controlling chlamydial spread is that the ...
    Measurement of the Promoter Activity in Escherichia coli by Using a Luciferase Reporter
    Authors:  Yuki Yamanaka, Hiroki Watanabe, Erika Yamauchi, Yukari Miyake and Kaneyoshi Yamamoto, date: 01/20/2020, view: 238, Q&A: 0
    [Abstract] The reporter system is widely used technique for measuring promoter activity in bacterial cells. Until now, a number of reporter system have been developed, but the bioluminescent reporter constructed from the bacterial luciferase genes is one of the useful systems for measuring in vivo dynamics of gene expression. The introduced ...
    Unbiased and Tailored CRISPR/Cas gRNA Libraries by Synthesizing Covalently-closed-circular (3Cs) DNA
    Authors:  Martin Wegner, Koraljka Husnjak and Manuel Kaulich, date: 01/05/2020, view: 701, Q&A: 0
    [Abstract] Simplicity, efficiency and versatility of the CRISPR/Cas system greatly contributed to its rapid use in a broad range of fields. Applications of unbiased CRISPR/Cas screenings are increasing and thus there is a growing need for unbiased and tailored CRISPR/Cas gRNA libraries. Conventional methods for gRNA library generation apply PCR and cloning ...
    High-throughput Site-directed Scanning Mutagenesis Using a Two-fragment PCR Approach
    Authors:  Franziska M. Heydenreich, Tamara Miljuš, Dalibor Milić and Dmitry B. Veprintsev, date: 01/05/2020, view: 397, Q&A: 0
    [Abstract] Site-directed scanning mutagenesis is a useful tool applied in studying protein function and designing proteins with new properties, such as increased stability or enzymatic activity. Creating a systematic library of hundreds of site-directed mutants is still a demanding and expensive task. The established protocols for making such libraries ...
    Localizing Genome Segments and Protein Products of a Multipartite Virus in Host Plant Cells
    Authors:  Marie-Stephanie Vernerey, Elodie Pirolles, Stephane Blanc and Anne Sicard, date: 12/05/2019, view: 531, Q&A: 0
    [Abstract] A founding paradigm in virology is that the spatial unit of the viral replication cycle is an individual cell. This concept applied to multipartite viruses–which have a genome composed of two or more nucleic acid segments, each individually encapsulated–implies that all segments constituting a viral genome need to coinfect the same host cell for ...
    Enhanced-ice-COLD-PCR for the Sensitive Detection of Rare DNA Methylation Patterns in Liquid Biopsies
    Authors:  Florence Mauger and Jörg Tost, date: 12/05/2019, view: 434, Q&A: 0
    [Abstract] In the context of precision medicine, the identification of novel biomarkers for the diagnosis of disease, prognosis, predicting treatment outcome and monitoring of treatment success is of great importance. The analysis of methylated circulating-cell free DNA provides great promise to complement or replace genetic markers for these applications, ...
    DNase I Chromatin Accessibility Analysis
    Authors:  Brook S. Nepon-Sixt and Mark G. Alexandrow, date: 12/05/2019, view: 428, Q&A: 0
    [Abstract] Chromatin consists of compacted DNA in complex with proteins and contributes to the organization of DNA and its stability. Furthermore, chromatin plays key roles in regulating cellular processes such as DNA replication, transcription, DNA repair, and mitosis. Chromatin assumes more compact (inaccessible) or decondensed (accessible) conformations ...
    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: 519, 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). ...
    Quantification of HIV-2 DNA in Whole Blood
    Authors:  Zsófia Szojka, Sara Karlson, Marianne Jansson and Patrik Medstrand, date: 10/20/2019, view: 676, 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 ...



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