分子生物学

分类

    现刊
    Direct-TRI: High-throughput RNA-extracting Method for all Stages of Zebrafish Development
    Direct-TRI: 斑马鱼发育各阶段高通量RNA提取方法
    作者:Kota Ujibe, Kanako Nishimura, Makoto Kashima and Hiromi Hirata日期:09/05/2021,浏览量:412,Q&A: 0

    Recent popularization of next-generation sequencing enables conducting easy transcriptome analysis. Nevertheless, substantial RNA isolation work prior to RNA sequencing, as well as the high cost involved, still makes the routine use of large-scale

    ...
    In vitro Cleavage and Electrophoretic Mobility Shift Assays for Very Fast CRISPR
    快速CRISPR的体外裂解和电泳迁移率位移测定
    作者:Roger S. Zou, Yang Liu and Taekjip Ha日期:09/05/2021,浏览量:605,Q&A: 0

    CRISPR-Cas9 has transformed biomedical research and medicine through convenient and targeted manipulation of DNA. Time- and spatially-resolved control over Cas9 activity through the recently developed very fast CRISPR (vfCRISPR) system have

    ...
    Measurement of LRRK2 Kinase Activity by Proximity Ligation Assay
    通过邻近连接测定法测量 LRRK2 激酶活性
    作者:Matthew T. Keeney, Eric K. Hoffman, J. Timothy Greenamyre and Roberto Di Maio日期:09/05/2021,浏览量:407,Q&A: 0

    Missense mutations in leucine rich-repeat kinase 2 (LRRK2) cause forms of familial Parkinson’s disease and have been linked to ‘idiopathic’ Parkinson’s disease. Assessment of LRRK2 kinase activity has been very challenging

    ...
    Preparation and Characterization of Internally Modified DNA Templates for Chemical Transcription Roadblocking
    用于化学转录障碍的内部修饰 DNA 模板的制备和特征分析
    作者:Eric J. Strobel日期:09/05/2021,浏览量:321,Q&A: 0

    Site-specific transcription arrest is the basis of emerging technologies that assess nascent RNA structure and function. Cotranscriptionally folded RNA can be displayed from an arrested RNA polymerase (RNAP) for biochemical manipulations by halting

    ...
    U2.3 Precursor Small Nuclear RNA in vitro Processing Assay
    U2.3前体小核 RNA体外加工试验
    作者:Chan Lin, Yujie Feng, Xueyan Peng, Jiaming Wu, Weili Wang and Yunfeng Liu日期:09/05/2021,浏览量:261,Q&A: 0

    Small nuclear RNAs (snRNAs) are vital for eukaryotic cell activities and play important roles in pre-mRNA splicing. The molecular mechanism underlying the transcription of snRNA, regulated via upstream/downstream cis-elements and relevant

    ...
    In situ Hybridization of miRNAs in Human Embryonic Kidney and Human Pluripotent Stem Cell-derived Kidney Organoids
    在人胚肾和人多能干细胞衍生的肾类器官中微小RNA的原位杂交
    作者:Filipa M. Lopes, Susan J. Kimber and Ioannis Bantounas日期:09/05/2021,浏览量:347,Q&A: 0

    MicroRNAs are small RNAs that negatively regulate gene expression and play an important role in fine-tuning molecular pathways during development. There is increasing interest in studying their function in the kidney, but the majority of studies to

    ...
    Kinetic Analysis of a Protein-protein Complex to Determine its Dissociation Constant (KD) and the Effective Concentration (EC50) of an Interplaying Effector Molecule Using Bio-layer Interferometry
    蛋白质-蛋白质复合物的动力学分析,以确定其电离常数 (KD) 和使用生物层干涉法相互作用的效应分子的有效浓度 (EC50)
    作者:Tim Orthwein, Luciano F. Huergo, Karl Forchhammer and Khaled A. Selim日期:09/05/2021,浏览量:897,Q&A: 0

    Biolayer interferometry (BLI) is an emerging analytical tool that allows the study of protein complexes in real time to determine protein complex kinetic parameters. This article describes a protocol to determine the KD of a protein complex using a

    ...
    Kinetic Analysis of a Protein-protein Complex to Determine its Dissociation Constant (KD) and the Effective Concentration (EC50) of an Interplaying Effector Molecule Using Bio-layer Interferometry
    蛋白质-蛋白质复合物的动力学分析,以确定其电离常数 (KD) 和使用生物层干涉法相互作用的效应分子的有效浓度 (EC50)
    作者:Tim Orthwein, Luciano F. Huergo, Karl Forchhammer and Khaled A. Selim日期:09/05/2021,浏览量:897,Q&A: 0
    [Abstract]

    Biolayer interferometry (BLI) is an emerging analytical tool that allows the study of protein complexes in real time to determine protein complex kinetic parameters. This article describes a protocol to determine the KD of a protein complex using a 6×His tagged fusion protein as bait immobilized on the NTA sensor chip of the

    ...
    In situ Hybridization of miRNAs in Human Embryonic Kidney and Human Pluripotent Stem Cell-derived Kidney Organoids
    在人胚肾和人多能干细胞衍生的肾类器官中微小RNA的原位杂交
    作者:Filipa M. Lopes, Susan J. Kimber and Ioannis Bantounas日期:09/05/2021,浏览量:347,Q&A: 0
    [Abstract]

    MicroRNAs are small RNAs that negatively regulate gene expression and play an important role in fine-tuning molecular pathways during development. There is increasing interest in studying their function in the kidney, but the majority of studies to date use kidney cell lines and assess the total amounts of miRNAs of interest either by qPCR or by

    ...
    U2.3 Precursor Small Nuclear RNA in vitro Processing Assay
    U2.3前体小核 RNA体外加工试验
    作者:Chan Lin, Yujie Feng, Xueyan Peng, Jiaming Wu, Weili Wang and Yunfeng Liu日期:09/05/2021,浏览量:261,Q&A: 0
    [Abstract]

    Small nuclear RNAs (snRNAs) are vital for eukaryotic cell activities and play important roles in pre-mRNA splicing. The molecular mechanism underlying the transcription of snRNA, regulated via upstream/downstream cis-elements and relevant trans-elements, has been investigated in detail using cell-free extracts. However, the processing of precursor

    ...
    Preparation and Characterization of Internally Modified DNA Templates for Chemical Transcription Roadblocking
    用于化学转录障碍的内部修饰 DNA 模板的制备和特征分析
    作者:Eric J. Strobel日期:09/05/2021,浏览量:321,Q&A: 0
    [Abstract]

    Site-specific transcription arrest is the basis of emerging technologies that assess nascent RNA structure and function. Cotranscriptionally folded RNA can be displayed from an arrested RNA polymerase (RNAP) for biochemical manipulations by halting transcription elongation at a defined DNA template position. Most transcription

    ...
    Measurement of LRRK2 Kinase Activity by Proximity Ligation Assay
    通过邻近连接测定法测量 LRRK2 激酶活性
    作者:Matthew T. Keeney, Eric K. Hoffman, J. Timothy Greenamyre and Roberto Di Maio日期:09/05/2021,浏览量:407,Q&A: 0
    [Abstract]

    Missense mutations in leucine rich-repeat kinase 2 (LRRK2) cause forms of familial Parkinson’s disease and have been linked to ‘idiopathic’ Parkinson’s disease. Assessment of LRRK2 kinase activity has been very challenging due to its size, complex structure, and relatively low abundance. A standard in the field to assess

    ...
    In vitro Cleavage and Electrophoretic Mobility Shift Assays for Very Fast CRISPR
    快速CRISPR的体外裂解和电泳迁移率位移测定
    作者:Roger S. Zou, Yang Liu and Taekjip Ha日期:09/05/2021,浏览量:605,Q&A: 0
    [Abstract]

    CRISPR-Cas9 has transformed biomedical research and medicine through convenient and targeted manipulation of DNA. Time- and spatially-resolved control over Cas9 activity through the recently developed very fast CRISPR (vfCRISPR) system have facilitated comprehensive studies of DNA damage and repair. Understanding the fundamental principles of Cas9

    ...
    Direct-TRI: High-throughput RNA-extracting Method for all Stages of Zebrafish Development
    Direct-TRI: 斑马鱼发育各阶段高通量RNA提取方法
    作者:Kota Ujibe, Kanako Nishimura, Makoto Kashima and Hiromi Hirata日期:09/05/2021,浏览量:412,Q&A: 0
    [Abstract]

    Recent popularization of next-generation sequencing enables conducting easy transcriptome analysis. Nevertheless, substantial RNA isolation work prior to RNA sequencing, as well as the high cost involved, still makes the routine use of large-scale transcriptome analysis difficult. For example, conventional phenol-chloroform RNA extraction cannot

    ...
    Optimised Method for the Production and Titration of Lentiviral Vectors Pseudotyped with the SARS-CoV-2 Spike
    SARS-CoV-2刺突慢病毒伪型载体制备和滴定的优化方法
    [Abstract]

    The use of recombinant lentivirus pseudotyped with the coronavirus Spike protein of SARS-CoV-2 would circumvent the requirement of biosafety-level 3 (BSL-3) containment facilities for the handling of SARS-CoV-2 viruses. Herein, we describe a fast and reliable protocol for the transient production of lentiviruses pseudotyped with SARS-CoV-2 Spike

    ...
    Implementing Novel Designs in pET Expression Plasmids that Increase Protein Production
    在pET表达质粒中实现增加蛋白质产量的新设计
    作者:Patrick J. Shilling and Daniel O. Daley日期:08/20/2021,浏览量:1275,Q&A: 0
    [Abstract]

    pET expression plasmids are widely used in the biotechnology, biopharmaceutical, and basic research sectors for the production of recombinant proteins. Typically, they are used off-the-shelf because they support high production titers; however, we have identified two design flaws in many pET plasmids that limit their production capacity. We used

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    Neutral Comet Assay to Detect and Quantitate DNA Double-Strand Breaks in Hematopoietic Stem Cells
    检测和定量造血干细胞DNA双链断裂的中性彗星试验
    作者:Irene Mariam Roy, Pon Sowbhagya Nadar and Satish Khurana日期:08/20/2021,浏览量:461,Q&A: 0
    [Abstract]

    In vertebrates, hematopoietic stem cells (HSCs) regulate the supply of blood cells throughout the lifetime and help to maintain homeostasis. Due to their long lifespan, genetic integrity is paramount for these cells, and accordingly, a number of stem cell-specific mechanisms are employed. However, HSCs tend to show more DNA damage with increasing

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