Molecular Biology

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    A Gel-Based Assay for Probing Protein Translocation on dsDNA
    Authors:  Christiane Brugger and Alexandra M. Deaconescu, date: 07/20/2021, view: 871, Q&A: 0

    Protein translocation on DNA represents the key biochemical activity of ssDNA translocases (aka helicases) and dsDNA translocases such as chromatin remodelers. Translocation depends on DNA binding but is a distinct process as it typically involves

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    Extraction of DNA and RNA from Formalin-fixed Paraffin-embedded Tissue Specimens

    One of the most common methods for isolating biomolecules from clinical samples is using TRIzol. While many tissues are freshly frozen, a large number of tissues are formalin-fixed paraffin-embedded (FFPE) blocks for pathology and diagnostics.

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    Modeling Perturbations in Protein Filaments at the Micro and Meso Scale Using NAMD and PTools/Heligeom
    Authors:  Benjamin Boyer, Benoist Laurent, Charles H. Robert and Chantal Prévost, date: 07/20/2021, view: 874, Q&A: 0

    Protein filaments are dynamic entities that respond to external stimuli by slightly or substantially modifying the internal binding geometries between successive protomers. This results in overall changes in the filament architecture, which are

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    GeneWeld: Efficient Targeted Integration Directed by Short Homology in Zebrafish

    Efficient precision genome engineering requires high frequency and specificity of integration at the genomic target site. Multiple design strategies for zebrafish gene targeting have previously been reported with widely varying frequencies for

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    Measurement of Transgene Copy Number in Plants Using Droplet Digital PCR
    Authors:  Yao-Min Cai, Quentin M. Dudley and Nicola J. Patron, date: 07/05/2021, view: 993, Q&A: 0

    Transgenic plants are produced both to investigate gene function and to confer desirable traits into crops. Transgene copy number is known to influence expression levels, and consequently, phenotypes. Similarly, knowledge of transgene zygosity is

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    PCR-mediated One-day Synthesis of Guide RNA for the CRISPR/Cas9 System
    Authors:  Naim Hassan, Farhana Easmin, Keisuke Ekino and Satoshi Harashima, date: 07/05/2021, view: 1101, Q&A: 0

    Nowadays, CRISPR (clustered regularly interspaced short palindromic repeats) and the CRISPR-associated protein (Cas9) system play a major role in genome editing. To target the desired sequence of the genome successfully, guide RNA (gRNA) is

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    Simplified Epigenome Profiling Using Antibody-tethered Tagmentation
    Authors:  Steven Henikoff, Jorja G. Henikoff and Kami Ahmad, date: 06/05/2021, view: 1261, Q&A: 0

    We previously introduced Cleavage Under Targets & Tagmentation (CUT&Tag), an epigenomic profiling method in which antibody tethering of the Tn5 transposase to a chromatin epitope of interest maps specific chromatin features in small samples

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    ATAC-Seq-based Identification of Extrachromosomal Circular DNA in Mammalian Cells and Its Validation Using Inverse PCR and FISH
    Authors:  Zhangli Su, Shekhar Saha, Teressa Paulsen, Pankaj Kumar and Anindya Dutta, date: 05/05/2021, view: 3122, Q&A: 0

    Recent studies from multiple labs including ours have demonstrated the importance of extrachromosomal circular DNA (eccDNA) from yeast to humans (Shibata et al., 2012; Dillon et al., 2015; Møller et al., 2016; Kumar et al., 2017; Turner et

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    A Sensitive and Specific PCR-based Assay to Quantify Hepatitis B Virus Covalently Closed Circular (ccc) DNA while Preserving Cellular DNA
    Authors:  Benno Zehnder, Stephan Urban and Thomas Tu, date: 04/20/2021, view: 2609, Q&A: 0

    Hepatitis B virus (HBV) is the major cause of liver diseases and liver cancer worldwide. After infecting hepatocytes, the virus establishes a stable episome (covalently closed circular DNA, or cccDNA) that serves as the template for all viral

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    A Novel Method to Construct Binary CRISPR Vectors for Plant Transformation by Single Round of PCR Amplification
    Authors:  Kang Li, Yuhui Wang and Chuanying Fang, date: 04/05/2021, view: 2452, Q&A: 0

    CRISPR/Cas9 is an established and flexible tool for genome editing. However, most methods used to generate expression clones for the CRISPR/Cas9 are time-consuming. Hence, we have developed a one-step protocol to introduce sgRNA expression

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