Titlebook: RNA Amplification and Analysis; Methods and Protocol Mekbib Astatke Book 2024 JHU Applied Physics Laboratory 2024 RNA extraction.multiplex

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RNA Analysis Using Immunoassay Detection Formath a pre-coated antibody (Ab) directed at the hapten moiety; (c) sandwich complex formation with an Ab that selectively recognizes the DNA/RNA structural motif; and (d) detection of the sandwich complex using a secondary Ab enzyme conjugate targeting the anti-DNA/RNA Ab followed by standard enzyme-linked immunosorbent assay (ELISA) visualization.

In-Situ

Sequencing: 10X Genomics 3′ HT Assay for Gene Expressionto 16 samples per run. The 3′ HT assay supports whole cells and nuclei as input, with an approximate 60% capture rate. Here we describe the methods for sample quality control (QC) assays, loading and operation of the Chromium X instrument for cell capture, and cDNA synthesis and library preparation for downstream Illumina sequencing.

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RNA Isolation from Human Stem Cell–Derived Retinal Organoidsicient quantity, and suitable quality, of RNA from very small amounts of tissue and/or samples containing low numbers of cells. Here we outline a total RNA isolation method that reproducibly yields high-quality RNA from human stem cell–derived retinal organoids for downstream transcriptomic analysis.

熒光

Single-Cell Imaging of mRNA by Target RNA-Initiated RCAplate, this method can efficiently detect mRNA in the absence of reverse transcription. This method enables spatial localization and correlation analysis of gene expression in single cells, which helps us to elucidate gene function and regulatory mechanisms.

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Assessing MicroRNA Profiles from Low Concentration Extracellular Vesicle RNA Utilizing NanoString nCbe a method to adapt NanoString nCounter microRNA profiling to assess mature microRNA expression in low-concentration RNA samples, including concentrating the RNA, quantifying the RNA, and performing the NanoString protocol. Twelve samples can be assessed at one time using this method.

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Direct Detection of SARS-CoV-2 RNA in Saliva with Colorimetric RT-LAMPmprehensive step-by-step protocol for establishing an RT-LAMP-based test for direct detection of SARS-CoV-2 genomic RNA in saliva samples using different colorimetric detection methods. Importantly, this versatile test can be easily adapted to detect emerging pathogens.

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RNA Isolation from Human Stem Cell–Derived Retinal OrganoidsCR), Northern blotting, microarrays, and RNA-sequencing. While many RNA purification methods have been reported, it can be challenging to extract sufficient quantity, and suitable quality, of RNA from very small amounts of tissue and/or samples containing low numbers of cells. Here we outline a tota