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    Preparation and Purification of Proteins Secreted from Phytophthora sojae
    Authors:  Yeqiang Xia, Yan Wang and Yuanchao Wang, date: 10/20/2018, view: 25, Q&A: 0
    Phytophthora sojae, the causal agent of soybean root and stem rot, is responsible for enormous economic losses in soybean production. P. sojae secrets various effectors to reprogram host immunity. The plant apoplastic space is a ...
    Microtitre Plate Based Cell-SELEX Method
    Authors:  Munish Shorie and Harmanjit Kaur, date: 10/20/2018, view: 27, Q&A: 0
    Aptamers have emerged as a novel category in the field of bioreceptors due to their wide applications ranging from biosensing to therapeutics. Several variations of their screening process, called SELEX have been reported which can yield sequences ...
    Detection and Differentiation of Multiple Viral RNAs Using Branched DNA FISH Coupled to Confocal Microscopy and Flow Cytometry
    Authors:  Nicholas van Buuren and Karla Kirkegaard, date: 10/20/2018, view: 29, Q&A: 0
    Due to the exceptionally high mutation rates of RNA-dependent RNA polymerases, infectious RNA viruses generate extensive sequence diversity, leading to some of the lowest barriers to the development of antiviral drug resistance in the microbial ...
    Behavioral Evaluation of Seeking and Preference of Alcohol in Mice Subjected to Stress
    Authors:  Ana Canseco-Alba, Norman Schanz, Hiroki Ishiguro, Qing-Rong Liu and Emmanuel S. Onaivi, date: 10/20/2018, view: 18, Q&A: 0
    The alcohol preference model is one of the most widely used animal models relevant to alcoholism. Stressors increase alcohol consumption. Here we present a protocol for a rapid and useful tool to test alcohol preference and stress-induced alcohol ...
    Generation of Gene Knockout and Gene Replacement with Complete Removal of Full-length Endogenous Transcript Using CRISPR-Trap
    Authors:  Jonas Mechtersheimer, Stefan Reber and Marc-David Ruepp, date: 10/20/2018, view: 21, Q&A: 0
    This protocol describes the application of the CRISPR-Trap from designing of the gene targeting strategy to validation of successfully edited clones that was validated on various human cell lines, among them human induced pluripotent stem cells ...
    Preparation and Purification of Proteins Secreted from Phytophthora sojae
    Authors:  Yeqiang Xia, Yan Wang and Yuanchao Wang, date: 10/20/2018, view: 25, Q&A: 0
    [Abstract] Phytophthora sojae, the causal agent of soybean root and stem rot, is responsible for enormous economic losses in soybean production. P. sojae secrets various effectors to reprogram host immunity. The plant apoplastic space is a major battleground in plant-pathogen interactions. Here we describe a protocol for purification and ...
    Microtitre Plate Based Cell-SELEX Method
    Authors:  Munish Shorie and Harmanjit Kaur, date: 10/20/2018, view: 27, Q&A: 0
    [Abstract] Aptamers have emerged as a novel category in the field of bioreceptors due to their wide applications ranging from biosensing to therapeutics. Several variations of their screening process, called SELEX have been reported which can yield sequences with desired properties needed for their final use. We report a facile microtiter plate-based ...
    Detection and Differentiation of Multiple Viral RNAs Using Branched DNA FISH Coupled to Confocal Microscopy and Flow Cytometry
    Authors:  Nicholas van Buuren and Karla Kirkegaard, date: 10/20/2018, view: 29, Q&A: 0
    [Abstract] Due to the exceptionally high mutation rates of RNA-dependent RNA polymerases, infectious RNA viruses generate extensive sequence diversity, leading to some of the lowest barriers to the development of antiviral drug resistance in the microbial world. We have previously discovered that higher barriers to the development of drug resistance can be ...
    Behavioral Evaluation of Seeking and Preference of Alcohol in Mice Subjected to Stress
    Authors:  Ana Canseco-Alba, Norman Schanz, Hiroki Ishiguro, Qing-Rong Liu and Emmanuel S. Onaivi, date: 10/20/2018, view: 18, Q&A: 0
    [Abstract] The alcohol preference model is one of the most widely used animal models relevant to alcoholism. Stressors increase alcohol consumption. Here we present a protocol for a rapid and useful tool to test alcohol preference and stress-induced alcohol consumption in mice. In this model, animals are given two bottles, one with a diluted solution of ...
    Generation of Gene Knockout and Gene Replacement with Complete Removal of Full-length Endogenous Transcript Using CRISPR-Trap
    Authors:  Jonas Mechtersheimer, Stefan Reber and Marc-David Ruepp, date: 10/20/2018, view: 21, Q&A: 0
    [Abstract] This protocol describes the application of the CRISPR-Trap from designing of the gene targeting strategy to validation of successfully edited clones that was validated on various human cell lines, among them human induced pluripotent stem cells (hiPSCs). The advantage of CRISPR-Trap over conventional approaches is the complete removal of any ...
    Real-time PCR Analysis of PAMP-induced Marker Gene Expression in Nicotiana benthamiana
    Authors:  Fan Liu, Yuanpeng Xu, Yan Wang and Yuanchao Wang, date: 10/05/2018, view: 156, Q&A: 0
    [Abstract] Perception of pathogen-associated molecular patterns (PAMPs) often triggers various innate immune responses in plants. The transcriptional changes of defense-related genes are often used as a marker to assay PAMP-triggered plant immune response. Here we described a protocol to monitor the relative expression level of marker genes in Nicotiana ...
    Preparation of Sequencing RNA Libraries through Chemical Cross-linking Coupled to Affinity Purification (cCLAP) in Saccharomyces cerevisiae
    Authors:  Congwei Wang, Julie Weidner and Anne Spang, date: 10/05/2018, view: 368, Q&A: 0
    [Abstract] Ribonucleoprotein particles (mRNPs) are complexes consisting of mRNAs and RNA-binding proteins (RBPs) which control mRNA transcription localization, turnover, and translation. Some mRNAs within the mRNPs have been shown to undergo degradation or storage. Those transcripts can lack general mRNA elements, like the poly(A) tail or 5’ cap structure, ...
    Fluorescence Titrations to Determine the Binding Affinity of Cyclic Nucleotides to SthK Ion Channels
    Authors:  Philipp A.M. Schmidpeter and Crina M. Nimigean, date: 10/05/2018, view: 104, Q&A: 0
    [Abstract] The cyclic-nucleotide modulated ion channel family includes cyclic nucleotide-gated (CNG) and hyperpolarization-activated and cyclic nucleotide-modulated (HCN) channels, which play essential roles in visual and olfactory signaling and the heart pacemaking activity. Functionally, these channels have been extensively characterized by ...
    CRISPR/Cas9-mediated ssDNA Recombineering in Corynebacterium glutamicum
    Authors:  Jiao Liu, Yu Wang, Ping Zheng and Jibin Sun, date: 10/05/2018, view: 108, Q&A: 0
    [Abstract] Corynebacterium glutamicum is a versatile workhorse for industrial bioproduction of many kinds of chemicals and fuels, notably amino acids. Development of advanced genetic engineering tools is urgently demanded for systems metabolic engineering of C. glutamicum. Recently unveiled clustered regularly interspaced short palindromic ...
    Identifying Protein Interactions with Histone Peptides Using Bio-layer Interferometry
    Authors:  Bingbing Ren, Ahmed Mahmoud Mohammed Sayed, Hwei Ling Tan, Yu Keung Mok and Ee Sin Chen, date: 09/20/2018, view: 361, Q&A: 0
    [Abstract] Histone post-translational modifications (PTMs) regulate numerous cellular processes, including gene transcription, cell division, and DNA damage repair. Most histone PTMs affect the recruitment or exclusion of reader proteins from chromatin. Here, we present a protocol to measure affinity and interaction kinetics between histone peptides and the ...