Protocols in Current Issue
    An Inexpensive and Comprehensive Method to Examine and Quantify Field Insect Community Influenced by Host Plant Olfactory Cues
    Authors:  Rupesh Kariyat, Jesus Chavana and Jasleen Kaur, date: 08/20/2018, view: 1345, Q&A: 0
    [Abstract] Insect pollinators, herbivores and their natural enemies use olfactory cues emitted by their host plants to locate them. In insect-plant ecology, understanding the mechanisms underlying these interactions are of critical importance, as this bio-communication has both ecological and agricultural applications. However, the first step in such ...
    Monitoring Xylem Hydraulic Pressure in Woody Plants
    [Abstract] Xylem sap circulates under either positive or negative hydraulic pressure in plants. Negative hydraulic pressure (i.e., tension) is the most common situation when transpiration is high, and several devices have been developed to quantify it accurately (e.g., Scholander pressure chamber, psychrometers). However, a proper ...
    Construction of a Single Transcriptional Unit for Expression of Cas9 and Single-guide RNAs for Genome Editing in Plants
    Authors:  Xu Tang, Zhaohui Zhong, Xuelian Zheng and Yong Zhang, date: 09/05/2017, view: 5665, Q&A: 0
    [Abstract] The CRISPR (clustered regularly interspaced short palindromic repeats)-associated protein9 (Cas9) is a simple and efficient tool for genome editing in many organisms including plant and crop species. The sgRNAs of the CRISPR/Cas9 system are typically expressed from RNA polymerase III promoters, such as U6 and U3. In many transformation events, ...
    Protein Isolation from Plasma Membrane, Digestion and Processing for Strong Cation Exchange Fractionation
    Authors:  Ning Yang, Bing Han and Tai Wang, date: 05/20/2017, view: 4683, Q&A: 0
    [Abstract] Plasma membrane (PM) proteins play crucial roles in diverse biological processes. But their low abundance, alkalinity and hydrophobicity make their isolation a difficult task. This protocol describes an efficient method for PM proteins isolation, digestion and fractionation so that they can be well prepared for mass spectrometry analysis.
    Ribosomal RNA N-glycosylase Activity Assay of Ribosome-inactivating Proteins
    Authors:  Rosario Iglesias, Lucía Citores and José M. Ferreras, date: 03/20/2017, view: 5358, Q&A: 0
    [Abstract] Ribosome-inactivating proteins (RIPs) are enzymes that irreversibly inactivate ribosomes as a consequence of their N-glycosylase (EC activity. The enzyme cleaves the N-glycosidic bond between the adenine No. 4324 from the 28S rRNA and its ribose in rat ribosomes (or the equivalent adenine in sensitive ribosomes from other organisms). ...
    Acetyl Bromide Soluble Lignin (ABSL) Assay for Total Lignin Quantification from Plant Biomass
    Authors:  William J. Barnes and Charles T. Anderson, date: 03/05/2017, view: 7160, Q&A: 0
    [Abstract] Lignin is the second most abundant biopolymer on Earth, providing plants with mechanical support in secondary cell walls and defense against abiotic and biotic stresses. However, lignin also acts as a barrier to biomass saccharification for biofuel generation (Carroll and Somerville, 2009; Zhao and Dixon, 2011; Wang et al., 2013). For ...
    PEA-CLARITY: Three Dimensional (3D) Molecular Imaging of Whole Plant Organs
    [Abstract] Here we report the adaptation of the CLARITY technique to plant tissues with addition of enzymatic degradation to improve optical clearing and facilitate antibody probe penetration. Plant-Enzyme-Assisted (PEA)-CLARITY, has allowed deep optical visualisation of stains, expressed fluorescent proteins and IgG-antibodies in tobacco and Arabidopsis ...
    Mungbean Yellow Mosaic India Virus (MYMIV)-infection, Small RNA Library Construction and Deep Sequencing for MicroRNA Identification in Vigna mungo
    Authors:  Anirban Kundu, Sujay Paul, Amita Pal and Genotypic Technology, date: 10/20/2016, view: 5653, Q&A: 0
    [Abstract] This protocol describes small RNA library preparation from Vigna mungo total RNA followed by deep sequencing and analysis for microRNA identification.​
    Cycloheximide Assays to Measure Protein Degradation in vivo in Plants
    Authors:  Jonathan Gilkerson, Raymond Tam, Aimee Zhang, Kate Dreher and Judy Callis, date: 09/05/2016, view: 8176, Q&A: 0
    [Abstract] The half-life of a protein is a characteristic property, and can be modulated by post-translational modifications, changes in subcellular localization, and/or interaction with other proteins or ligands. As one determinant of its steady-state level, a protein’s degradation represents an important distinguishing attribute relevant to its biological ...
    Experimental Pipeline for SNP and SSR Discovery and Genotyping Analysis of Mango (Mangifera indica L.)
    [Abstract] Establishing a reservoir of polymorphic markers is an important key for marker-assisted breeding. Many crops are still lack of such genomic infrastructure. Single nucleotide polymorphisms (SNPs) and simple sequence repeats (SSRs) are useful as markers because they are widespread over the genome and many technologies were developed for high ...

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