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Protocols

protocols
Protocol

Plasmid Purification And Troubleshooting

Experiment Tags: Plasmid purification, Plasmid purification

Experiment Summary: Plasmids are small circular deoxyribonucleic acids (DNA) outside the chromosomes that replicate independently of chromosomal DNA. Although budding yeast and fission yeast can retain plasmids, the host of plasmids is almost always bacteria. This small circular DNA is widely used as a DNA vector in the fields of molecular biology, biochemistry, biotechnology, cell biology, etc.

protocols
Protocol

Comprehensive Guide to Stable Lentiviral Cell Line Construction

Experiment Tags: Lentiviral Vectors, Stable Cell Line, Gene Expression, Viral Transduction, Cell Culture, Puromycin Selection, Virus Packaging, Mammalian Cells, Transgene Expression

Experiment Summary: This protocol details the generation of stable cell lines using lentiviral vectors. The method covers plasmid preparation, 293T cell transfection for viral packaging, PEG8000-based virus concentration, target cell infection, antibiotic selection parameters, and single-clone isolation. Includes MOI calculation guidance, troubleshooting for common challenges like low viral titer and poor infection efficiency, and validation strategies for ensuring stable transgene expression.

protocols
Protocol

Bacterial Transformation Protocol

Experiment Tags: Bacterial Transformation, Bacterial Transformation Protocol, transformation, bacterial cells

Experiment Summary: Bacterial transformation is a fundamental process in molecular biology, enabling the introduction of foreign DNA into bacterial cells. Primarily used for the amplification and storage of plasmids, bacterial transformation is critical for cloning experiments.

protocols
Protocol

Protocol for Cell Viability Assays: CCK-8 and MTT

Experiment Tags: CCK-8 assay, MTT assay, cell viability, drug treatment, IC50, absorbance measurement, cell seeding, aseptic technique, OD values, 96-well plate, biological assay, cytotoxicity, experimental protocol.

Experiment Summary: This protocol provides a step-by-step guide for performing CCK-8 and MTT cell viability assays, covering cell seeding, drug concentration settings, treatment duration, and absorbance measurement. It also includes important considerations for avoiding contamination, ensuring cell health, and interpreting results accurately, especially in high-throughput drug testing.

protocols
Protocol

Monkeypox Antigen Expression and Antibody Response Protocol

Experiment Tags: MPXV, A29L protein, B6R protein, M1R protein, HEK293T cells, Recombinant proteins, Protein purification, SDS-PAGE, Western blot, ELISA, Neutralization assay, Antibody titer

Experiment Summary: This protocol outlines the process for expressing, purifying, and analyzing MPXV antigens and assessing their immunogenicity, which is vital for vaccine research and development.

protocols
Protocol

Dual-Fluorescence Flow Cytometry for Protein Level Analysis

Experiment Tags: Dual-fluorescence, Flow Cytometry, Protein Analysis, Steady-State Protein Levels, eGFP mCherry, Transfection Efficiency, Cell Line Transfection, Protein Abundance

Experiment Summary: This protocol provides a powerful tool for studying protein expression levels under various cellular conditions, offering single-cell resolution and the ability to detect subtle differences that might be masked in bulk assays. Its versatility makes it applicable to a wide range of proteins and cellular contexts, contributing to our understanding of protein homeostasis and cellular biology.

protocols
Protocol

CRISPR/Cas9-Mediated Gene Editing in C. elegans

Experiment Tags: CRISPR-Cas9, Gene Editing, C. elegans, sgRNA, Plasmid Construction Homologous Recombination, Genetic Screening.

Experiment Summary: This protocol outlines the process for performing CRISPR/Cas9-mediated genome modifications in C. elegans, including the design of specific sgRNA sequences, construction of Cas9-sgRNA plasmids, and preparation of injection mixtures.

protocols
Protocol

3D Tumorsphere Culture Protocol for Cancer Research

Experiment Tags: 3D Tumorsphere Culture, Cancer Research, Matrigel Embedding, Tumor Tissue Culture, Cell Transduction, Cryopreservation, Cell Counting

Experiment Summary: This protocol outlines a comprehensive procedure for 3D tumorsphere cultivation, including cell transduction, 3D tumorsphere culture using Matrigel, primary tumorsphere culture from fresh tumor tissue, spheroid passaging, and cryopreservation. The protocol aims to replicate the 3D environment of solid tumors, aiding researchers in studying tumor biology and conducting drug screening.

protocols
Protocol

Rapid and Efficient Gene Editing in Human Pluripotent Stem Cells via FLPe Recombinase-Mediated Cassette Exchange at the AAVS1 Locus

Experiment Tags: Gene Editing, FLPe Recombinase, Cassette Exchange, AAVS1 Locus, Human Pluripotent Stem Cells

Experiment Summary: The protocol presents a unique approach to FLPe recombinase-mediated cassette exchange (RMCE) gene editing at the AAVS1 locus in human pluripotent stem cells (hPSCs) that is both rapid and efficient. This method makes it easier to generate highly reliable isogenic lines, which in turn makes transgenesis studies in hPSCs easier to conduct.

protocols
Protocol

Protocol for CRISPR/Cas9-mediated Knockout in Difficult-to-Transfect Cells

Experiment Tags: CRISPR/Cas9, Lentivirus, Gene Knockout, Cell Line Development

Experiment Summary: The protocol utilizes lentiviral vectors and a control system to regulate Cas9 activity, enabling the development of knockout cell lines. The development of a human muscle cell line that targets the gene serves as evidence of the protocol's effectiveness.

* For research use only. Not intended for any clinical use.
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