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Experiment Tags: CRISPR-Cas9, Off-Target Activity, Chromosomal Translocation, LAM-HTGTS, iHTGTS
Experiment Summary: In this article, we described an improved HTGTS (iHTGTS) method, in which size-selection beads were used to enhance reaction efficiency and a new primer system was designed to be compatible with Illumina Hiseq sequencing. iHTGTS is a powerful method for comprehensively assessing Cas9 off-target effect.
Experiment Tags: Diagnosis, RNA Virus, Replicative Intermediate, dsRNA Isolation, dsRNA-Binding Protein
Experiment Summary: In this protocol, we describe a method for dsRNA isolation. Using this method, we can readily isolate viral dsRNA from a small amount of plant material, and can process numerous samples simultaneously.
Experiment Tags: Single Guide RNA Library, Library Design and Construction
Experiment Summary: This protocol describes how to generate a single guide RNA (sgRNA) library for use in genetic screening.
Experiment Tags: CircRNAs, RNA Pull-Down, RNA-Protein Interactions
Experiment Summary: In this protocol, we introduce a method to study RNA-protein interactions, which will facilitate researchers to carry out circRNA related experiments.
Experiment Tags: RSV Transcription, Minigenome, Polymerase, Luciferase Assay, Real-Time Quantitative PCR Assay
Experiment Summary: In this protocol, we detail two different methods of detecting viral RNA synthesis: a firefly luciferase assay for rapid and sensitive measurement of RSV polymerase activity; and a real-time quantitative PCR method for determination of specific effects on the transcription of individual viral genes and the polar transcription gradient of RSV.
Experiment Tags: CRISPRCas9 Library Screening, CRISPRCas9
Experiment Summary: In this protocol, we briefly describe the CRISPR/Cas9 library screening process to simplify and facilitate high-throughput screening.
Experiment Tags: TAC-Seq, Chromatin Accessibility, Genome Wide, Zebrafish, Tn5 Transposase
Experiment Summary: In this article, we describe the application of ATAC-seq in zebrafish embryos, thus illustrating the applicability of this method to the entire vertebrate population.
Experiment Tags: Adeno-Associated Virus, Binding, Internalization, Virus Entry, Virus, Cell Surface
Experiment Summary: In this protocol, we describe a quantitative PCR (qPCR) based method to determine the amount of vector bound to the cell surface and the amount of subsequent virus internalization based on viral genome quantification.
Experiment Tags: Circular RNA, In Vitro Transcription, Ligase, Phosphorylation, Polyacrylamide Gel Electrophoresis
Experiment Summary: This protocol describes a simple and straightforward way to obtain single-stranded circular RNA sequences in vitro.
Experiment Tags: CircRNAs, CircRNA Pulldown, Antisense Oligo, Biotin Pulldown, RT-qPCR, miRNA, RNA-Binding Proteins
Experiment Summary: In this detailed protocol, the circRNA of interest (e.g., circHipk2) was captured using a biotin-labeled antisense oligo (ASO) targeting the circHipk2 backsplice junction sequence followed by pulldown with streptavidin-conjugated magnetic beads. The specific enrichment of circRNA was analyzed using reverse transcription quantitative PCR (RT-qPCR).
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