Protocols in Current Issue
    Isolation and Transcriptomic Profiling of Single Myofibers from Mice
    [Abstract] Skeletal muscle is composed of different cells and myofiber types, with distinct metabolic and structural features. Generally, transcriptomic analysis of skeletal muscle is performed using whole muscle, resulting in average information as all cells composing the organ contribute to the expression value detected for each gene with the loss of ...
    Quantitative Live-cell Reporter Assay for Noncanonical Wnt Activity
    Authors:  Edith P. Karuna, Michael W. Susman and Hsin-Yi Henry Ho, date: 03/20/2018, view: 7643, Q&A: 0
    [Abstract] Noncanonical Wnt signaling functions independently of the β-catenin pathway to control diverse developmental processes, and dysfunction of the pathway contributes to a number of human pathological conditions, including birth defects and metastatic cancer. Progress in the field, however, has been hampered by the scarcity of functional assays for ...
    Identification of RNA-binding Proteins
    Authors:  Kazuya Masuda and Tadamitsu Kishimoto, date: 09/05/2016, view: 12443, Q&A: 0
    [Abstract] This protocol describes the extraction of RNA-binding proteins (RBPs) from cell lysates. In order to pull down target RBPs, 5-bromo-UTP (BrUTP)-incorporated RNA probes are used, which are generated by in vitro transcription. The schematic diagram (Flowchart) with procedure is indicated (Figure1 and Figure 2).

    Measurement of mRNA Decay in Mouse Embryonic Fibroblasts
    Authors:  Lian-Qun Qiu, Wi S. Lai, Deborah J. Stumpo and Perry J. Blackshear, date: 07/05/2016, view: 10287, Q&A: 0
    [Abstract] mRNA stability control is a critical step in the post-transcriptional regulation of gene expression. Actinomycin D, an antibiotic initially used as an anti-cancer drug, has turned out to be a convenient tool for studying the turnover rates of transcripts in cells, due to its inhibition of mRNA synthesis. Here, we describe a protocol for the ...
    Cellular Translational Reporter Assay
    Authors:  Taishi Kimura and Kiyoshi Takeda, date: 03/20/2014, view: 9974, Q&A: 0
    [Abstract] The method described here allows measuring the effect of exogenously introduced modifications to in vitro-transcribed mRNA on the translation in cells. Using cells derived from knockout mice and control littermates, this method enables to compare the results in the presence or absence of specific gene products. In our lab, we used this ...
    Single-cell Gene Expression Profiling of Mouse Stem Cells With Fluidigm BiomarkTM Dynamic Array
    Author:  Ana Sevilla, date: 05/05/2013, view: 17491, Q&A: 3
    [Abstract] This protocol describes how to use Fluidigm BiomarkTM 96.96 dynamic arrays for high-throughput expression profiling from single mouse stem cells, assaying up to 96 independent samples with up to 96 quantitative PCR (qPCR) probes (equivalent to 9,216 reactions) in a single experiment. This Dynamic Array contains a network of microfluidic ...

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