Titlebook: RNA Abundance Analysis; Methods and Protocol Hailing Jin,Isgouhi Kaloshian Book 2021Latest edition Springer Science+Business Media, LLC, pa

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CRISPR-Cas RNA Targeting Using Transient Cas13a Expression in ,iviral defense. We cover all the necessary information for cloning the Cas13 protein, crRNA guide cassette, performing transient .-mediated expression of the necessary Cas13a components and target RNA-virus, visualization of virus infection, and molecular quantification of viral accumulation using quantitative PCR.

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Strand-Specific RNA-Seq Applied to Malaria Samples,this method, we prepared high-quality strand-specific RNA-seq libraries from RNA extracted from the human malaria parasite . The libraries are compatible with Illumina.’s sequencers Genome Analyzer and Hi-Seq. The method can however be easily adapted to other platforms.

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Isolation of Insect Bacteriocytes as a Platform for Transcriptomic Analyses,its use. This chapter provides a simple, step-by-step dissection protocol for the rapid isolation of aphid bacteriocytes. We then describe an adapted protocol for efficient extraction and purification of bacteriocyte RNA that can be used for most downstream transcriptomic analyses.

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Small RNA Extraction and Quantification of Isolated Fungal Cells from Plant Tissue by the Sequentiahod). The isolated fungal cells are free of contaminants from the host plants, and remain viable, providing high-quality RNA for library construction. This method can be modified to isolate the infection structures of many other plant pathogens from plant tissue.

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Laser Microdissection of Cells and Isolation of High-Quality RNA After Cryosectioning,e, a protocol that combines good morphology preservation with RNA integrity maintenance was developed, and successfully applied to Arabidopsis and tomato galls. Specifically, early developing GCs at 3 and 7?days post-infection (dpi) were analyzed; RNA from LCM GCs was amplified and used successfully for microarray assays.

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,Analysis of RBP Regulation and Co-regulation of mRNA 3′ UTR Regions in a Luciferase Reporter SystemR), thus examining gene expression regulation on the mRNA level. Assessment of 3′ UTR sequence requirements, as well as single and co-regulatory roles of RBPs in regulation of mRNAs will be discussed.

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