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Molecular Biology

protocols
Protocol

Test MicroRNA Target with in vitro Cell Culture Luciferase Assay Protocol

Experiment Tags: MicroRNA, 3'UTR (3’UTR), Luciferase Assay, In Vitro

Experiment Summary: In this protocol, we present a protocol for the subcellular fractionation of cells to detect circRNA via RT-qPCR with divergent primers. Moreover, detailed steps for the generation of specific circRNAs-enriched cDNA included in this protocol will enhance the amplification and detection of low-abundance circRNAs.

protocols
Protocol

Antisense Oligo Pulldown of Circular RNA for Downstream Analysis Protocol

Experiment Tags: CircRNAs, CircRNA Pulldown, Antisense Oligo, Biotin Pulldown, RT-qPCR, miRNA, RNA-Binding Proteins

Experiment Summary: In this protocol, we describe protocols for the generation and enrichment of recombinant vaccinia viruses using targeted Cas9 as a selection tool. This novel use of Cas9 is a simple addition to current homologous recombination-based methods that are widespread in the field, facilitating implementation in laboratories already working with poxviruses.

protocols
Protocol

Preparation of Circular RNA In Vitro Protocol

Experiment Tags: Circular RNA, In Vitro Transcription, Ligase, Phosphorylation, Polyacrylamide Gel Electrophoresis

Experiment Summary: In this protocol, we describe in detail the necessary steps to deliver mRNA-LNP to the murine liver and, following intravenous injection of eGFP mRNA-LNP, verify transfection efficiency using flow cytometry and liver cell specificity using immunofluorescence analyses.

protocols
Protocol

Virus Binding and Internalization Protocol for Adeno-associated Virus

Experiment Tags: Adeno-Associated Virus, Binding, Internalization, Virus Entry, Virus, Cell Surface

Experiment Summary: In this protocol, we accurately describe specific assays that can deterministically detect anti-chikungunya virus antibodies (Ab) in the tissues or sera of infected mice.

protocols
Protocol

Assay for Transposase-Accessible Chromatin with High-Throughput Sequencing (ATAC-Seq) Protocol for Zebrafish Embryos

Experiment Tags: TAC-Seq, Chromatin Accessibility, Genome Wide, Zebrafish, Tn5 Transposase

Experiment Summary: In this protocol, we describe basic experimental procedures to produce ZIKV stocks and to quantify their concentration in infectious virus particles as well as to image and study this pathogen within infected cells using confocal microscopy-based imaging.