Efficient and Rapid Analysis of Polysomes and Ribosomal Subunits in Cells and Tissues Using Ribo Mega-SEC
    利用Ribo Mega-SEC高效快速分析细胞和组织中的多聚体和核糖体亚基
    作者:Harunori Yoshikawa, Ramasubramanian Sundaramoorthy, Daniel Mariyappa, Hao Jiang and Angus I. Lamond日期:08/05/2021,浏览量:1214,Q&A: 0

    Polysome profile analysis is a popular method for separating polysomes and ribosomal subunits and is typically achieved using a sucrose density gradient (SDG). This has remained the gold standard method since ribosomes were first discovered; however, this method is time-consuming and requires multiple steps from making the gradient and long

    A Detailed Protocol for Preparing Millimeter-sized Supergiant Liposomes that Permit Efficient Eukaryotic Cell-free Translation in the Interior
    作者:Hajime Takahashi and Atsushi Ogawa日期:06/20/2021,浏览量:880,Q&A: 0

    Liposomes have been used as a pseudo cell membrane for encapsulating biomolecules and creating an artificial cell in the interior where biochemical reactions can occur. Among the several methods used to prepare biomolecule-encapsulating liposomes, the spontaneous emulsion transfer method is superior to others in that it allows us to readily

    Spin Labeling of RNA Using “Click” Chemistry for Coarse-grained Structure Determination via Pulsed Electron-electron Double Resonance Spectroscopy
    作者:Maria F. Vicino, Tobias Hett and Olav Schiemann日期:05/05/2021,浏览量:2375,Q&A: 0

    Understanding the function of oligonucleotides on a molecular level requires methods for studying their structure, conformational changes, and internal dynamics. Various biophysical methods exist to achieve this, including the whole toolbox of Electron Paramagnetic Resonance (EPR or ESR) spectroscopy. An EPR method widely used in this regard is

    Monitoring Real-time Temperature Dynamics of a Short RNA Hairpin Using Förster Resonance Energy Transfer and Circular Dichroism
    作者:Martin Balcerowicz, Marco Di Antonio and Betty Y. W. Chung日期:03/20/2021,浏览量:2372,Q&A: 0

    RNA secondary structures are highly dynamic and subject to prompt changes in response to the environment. Temperature in particular has a strong impact on RNA structural conformation, and temperature-sensitive RNA hairpin structures have been exploited by multiple organisms to modify the rate of translation in response to temperature changes.

    Trypanosomatid, fluorescence-based in vitro U-insertion/U-deletion RNA-editing (FIDE)
    作者:Wolf-Matthias Leeder, Elisabeth Kruse and H. Ulrich Göringer日期:03/05/2021,浏览量:1781,Q&A: 0

    Gene expression within the mitochondria of African trypanosomes and other protozoan organisms relies on a nucleotide-specific RNA-editing reaction. In the process exclusively uridine (U)-nucleotides are site-specifically inserted into and deleted from sequence-deficient primary transcripts to convert them into translatable mRNAs. The reaction is

    Identification of Intrinsic RNA Binding Specificity of Purified Proteins by in vitro RNA Immunoprecipitation (vitRIP)
    作者:Marisa Müller, Tamas Schauer and Peter B. Becker日期:03/05/2021,浏览量:1522,Q&A: 0

    RNA-protein interactions are often mediated by dedicated canonical RNA binding domains. However, interactions through non-canonical domains with unknown specificity are increasingly observed, raising the question how RNA targets are recognized. Knowledge of the intrinsic RNA binding specificity contributes to the understanding of target

    Plant ARGONAUTE Protein Immunopurification for Pathogen Cross Kingdom Small RNA Analysis
    作者:Florian Dunker, Bernhard Lederer and Arne Weiberg日期:02/05/2021,浏览量:2100,Q&A: 0

    Over the last decade, it has been noticed that microbial pathogens and pests deliver small RNA (sRNA) effectors into their host plants to manipulate plant physiology and immunity for infection, known as cross kingdom RNA interference. In this process, fungal and oomycete parasite sRNAs hijack the plant ARGONAUTE (AGO)/RNA-induced silencing complex

    Ribosome Purification from an α-proteobacterium and rRNA Analysis by Northern Blot
    作者:Liang Yin and Caroline S. Harwood日期:12/05/2020,浏览量:1036,Q&A: 0
    [Abstract] Ribosomes are an integral part of cellular life. They are complex molecular machines consisting of multiple ribosomal proteins and RNAs. To study different aspects of ribosome composition, many methods have been developed over the decades. Here, we describe how to purify ribosomes from the α-proteobacterium Rhodopseudomonas palustris ...
    Fluorescent Polysome Profiling in Caenorhabditis elegans
    作者:Dan Shaffer and Jarod A Rollins日期:09/05/2020,浏览量:2666,Q&A: 0
    [Abstract] An important but often overlooked aspect of gene regulation occurs at the level of protein translation. Many genes are regulated not only by transcription but by their propensity to be recruited to actively translating ribosomes (polysomes). Polysome profiling allows for the separation of unbound 40S and 60S subunits, 80S monosomes, and actively ...
    RNA Stability Measurements Using RT-qPCR in Arabidopsis Seedlings
    作者:Tianran Jia and Brandon H. Le日期:07/20/2020,浏览量:2138,Q&A: 0
    [Abstract] Steady-state mRNA levels are determined by both the rates of transcription and degradation. Regulation of mRNA stability and/or degradation are key factors that can significantly affect mRNA levels and its biological functions. mRNA stability can be measured indirectly after transcription inhibition. This protocol described a rapid and sensitive ...