Protocols - Creative Biogene

Protocol

Protocol of Transient yet Robust Expression of Proteins in the Mouse Liver via Intravenous Injection of Lipid Nanoparticle-encapsulated Nucleoside-modified mRNA

Experiment Tags: Nucleoside-Modified mRNA, Lipid Nanoparticle, Liver Regeneration, Protein Expression, Retro-Orbital Injection

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

Protocol

Protocol for Analyzing of the Effect of Sphingomyelinase on Rubella Virus Infectivity in Two Cell Lines

Experiment Tags: Rubella Virus, Lipid, Sphingomyelin, Virus Entry, Sphingomyelinase

Experiment Summary: This protocol evaluates in detail the effect of SMase treatment on RuV infectivity in RK13 and Vero cells.

Protocol

Improved HTGTS for CRISPR/Cas9 Off-target Detection Protocol

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.

Protocol

A New Method to Isolate Total dsRNA

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.

Protocol

Single Guide RNA Library Design and Construction Protocol

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.

Protocol

RNA Pull-Down Protocol of CircRNA Research

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.

Protocol

Protocol for Studying RSV Transcription using Minigenome Systems

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.

Protocol

CRISPRCas9 Library Screening Protocol

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.

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 article, we describe the application of ATAC-seq in zebrafish embryos, thus illustrating the applicability of this method to the entire vertebrate population.

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 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.