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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).
Experiment Tags: MicroRNA, 3'UTR (3’UTR), Luciferase Assay, In Vitro
Experiment Summary: This protocol is to be used to test the binding and activity of miRNA on putative UTR target sequences. It is based on the expression of Luciferase as a reporter gene fused to the UTR sequence in the presence of plasmids containing pre-miRNA of interest or synthetic miRNA to test in an in vitro cell culture assay.
Experiment Tags: Glycine Oxidase Mutant Libraries, Random Mutagenesis
Experiment Summary: This protocol describes the construction of a glycine oxidase mutant library by random mutagenesis and the use of a phage T7 lysis-based approach for high-throughput screening of oxidase activity, among other things.
Experiment Tags: Long Noncoding RNAs, RNA-Protein Interaction, Mass Spectrometry
Experiment Summary: This article provides a detailed protocol for the assay with hands-on tips for the LnRNA pulldown assay, which is commonly used to identify protein partners of the cellular environment that interact with InRNA.
Experiment Tags: Glioma, Retrovirus, Intracerebral Injection, Murine Model, PDGFBB(PDGFBB), PTEN(PTEN), P53(P53)
Experiment Summary: This protocol describes a simple and practical method for inducing GBM formation by retrovirus injection in the murine brain. This system gives a spatial and temporal control over the induction of glioma and allows the assessment of therapeutic effects with a bioluminescent reporter.
Experiment Tags: Base-Editing, CRISPR-Cas9
Experiment Summary: This article provides optimized protocols (including experimental design, methods, and analyses) for designing base editing constructs for basic research.
Experiment Tags: Adeno-Associated Virus (AAV), PEG Aqua Partitioning, AAV Viral Purification, Pancreatic Ductal Infusion, Pancreatic Cell Lineage Tracing
Experiment Summary: This protocol describes efficient delivery of AAV to mediate GFP or Cre expression for cell lineage tracing in the mouse pancreas or for the delivery of transgenes under a specific promoter to these cells.
Experiment Tags: Single Cell, Genomics, Molecular Biology, Gene Expression, CRISPR
Experiment Summary: This protocol describes the detailed steps for generating knockout cell lines (coding or non-coding regions) using CRISPR/Cas9 tools.
Experiment Tags: Inflammation, Peritonitis, Sepsis, In Vivo Imaging, Myeloperoxidase
Experiment Summary: This protocol allows studying of inflammatory response induced by LPS that affect neutrophil infiltration by tracking myeloperoxidase (MPO) activity in vivo.
