Microbiology

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    Protocols in Current Issue
    An in vitro Assay of mRNA 3’ end Using the E. coli Cell-free Expression System
    Authors:  Monford Paul Abishek N and Heon M. Lim, date: 02/20/2022, view: 1251, Q&A: 0
    [Abstract]

    At the end of about 80% of the operon in Escherichia coli, translation termination decouples transcription, leading to Rho-dependent transcription termination (RDT). However, no in vitro or in vivo assay system has proven to be good enough to see the 3’ end of the mRNA generated by RDT. Here, we present a cell-free assay system that

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    Transcriptional Run-on: Measuring Nascent Transcription at Specific Genomic Sites in Yeast
    Authors:  Victoria Begley, Lola de Miguel-Jiménez and Sebastián Chávez, date: 06/20/2021, view: 1528, Q&A: 0
    [Abstract]

    DNA transcription by RNA polymerases has always interested the scientific community as it is one of the most important processes involved in genome expression. This has led scientists to come up with different protocols allowing analysis of this process in specific locations across the genome by quantitating the amount of RNA polymerases

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    In vitro Transcription (IVT) and tRNA Binding Assay
    Author:  Sonja MK Schoenfelder, date: 09/20/2014, view: 13382, Q&A: 0
    [Abstract] This protocol describes the coupling of (i) “live” in vitro RNA transcription with (ii) binding by a radiolabeled, pre-formed tRNA followed by native gel electrophoresis and phosphorimager scan to visualize the complex. The necessity arose from the stable structure that one RNA forms in the absence of its interaction partner. The T-box ...



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