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Protocol
Characterization of SARS-CoV-2 Glycoprotein Using a Quantitative Cell–Cell Fusion System
Coronaviruses (CoVs) infect host cells through the fusion of viral and cellular membrane and may also spread to the neighboring uninfected cells from infected cells through cell–cell fusion. The viral spike (S...
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Protocol
In Vivo and In Vitro Assay to Address Dendritic Cell Antigen Cross-Presenting Capacity
Antigen cross-presentation by dendritic cells is an important pathway to prime CD8+ T cells in infections, cancer, and other immune-mediated pathologies. Particularly in cancer, cross-presentation of tumor-associ...
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Protocol
Assessment of Dynein-Mediated Nuclear Migration in the Develo** Cortex by Live-Tissue Microscopy
During development of the cerebral cortex, neuroepithelial and radial glial cells undergo an oscillatory nuclear movement throughout their cell cycle, termed interkinetic nuclear migration. The nucleus of post...
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Protocol
Large-Scale Complete Sequencing and Haploty** of 1–10 kb DNA Molecules Using Short Massively Parallel Reads
In this chapter, we describe a simple, low-cost method for making many copies of a single DNA molecule (1–10 kb in length) as a concatemer on a long DNA strand. This can enable applications requiring high-qual...
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Protocol
A Simple Cost-Effective Method for Whole-Genome Sequencing, Haploty**, and Assembly
In this chapter, we describe single-tube long fragment read (stLFR), a simple preparation method for whole-genome sequencing and physical haploty** based on the DNA co-barcoding strategy. Similar to LFR, stL...
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Protocol
A Tutorial of EDTA: Extensive De Novo TE Annotator
Transposable elements (TEs) are important contributors to genome structure and evolution. With the growth of sequencing technologies, various computational pipelines and software programs have been developed t...
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Protocol
Cell-Derived Nanovesicles as Exosome-Mimetics for Drug Delivery Purposes: Uses and Recommendations
Cell-derived Drug Delivery Systems (DDSs), particularly exosomes, have grown in popularity and have been increasingly explored as novel DDSs, due to their intrinsic targeting capabilities. However, clinical tr...
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Protocol
Bead-Based Flow-Cytometric Cell Counting of Live and Dead Bacteria
Flow cytometry (FCM) is based on the detection of scattered light and fluorescence to identify cells with characteristics of interest. Many flow cytometers cannot precisely control the flow through its interro...
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Protocol
Biolistic Labeling of Retinal Ganglion Cells
Labeling of cellular structures is of fundamental importance in the investigation of diseases of the central nervous system. Biolistic labeling of retinal ganglion cells (RGCs) allows visualization of dendriti...
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Protocol
Migration of Q Cells in Caenorhabditis elegans
During C. elegans larval development, the Q neuroblasts produce their lineage by three rounds of divisions along with continuous cell migrations. Their neuronal progeny is dispersed from the pharynx to the anus. ...
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Protocol
Partial Hepatectomy and Castration of HBV Transgenic Mice
Hepatitis B virus (HBV) is a hepatotropic virus. Its infection can cause liver injury and regeneration, and its replication is affected by the gender. Transgenic mice that carry a 1.3-mer overlength HBV DNA ge...
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Protocol
Microarray Technology for Analysis of MicroRNA Expression in Renal Biopsies of Lupus Nephritis Patients
Systemic lupus erythematosus (SLE) is a complex autoimmune disease, which predominantly occurs in females and is characterized by autoantibody production against a host of nuclear self-antigens and deposition ...
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Protocol
Evaluating a Particular Circulating MicroRNA Species from an SLE Patient Using Stem-Loop qRT-PCR
Systemic lupus erythematosus (SLE) is a complex autoimmune disease, and correct judgment of SLE activity is very important in guiding precise clinical treatment. Circulating microRNAs (miRNAs) could serve as p...
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Protocol
Microfluidic-Based Manufacture of siRNA-Lipid Nanoparticles for Therapeutic Applications
A simple, efficient, and scalable manufacturing technique is required for develo** siRNA-lipid nanoparticles (siRNA-LNP) for therapeutic applications. In this chapter we describe a novel microfluidic-based m...
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Protocol
Polysaccharide-Derived Chiral Stationary Phases in Capillary Electrochromatography Enantioseparations
Capillary electrochromatography (CEC) has been shown to be a promising technique for miniaturized enantioseparations. Here we present the preparation and application of positively charged cellulose derivative-...
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Protocol
Microchip UV Absorbance Detection Applied to Isoelectric Focusing of Proteins
Isoelectric focusing (IEF) is considered as an attractive separation technique for biologically amphoteric compounds (e.g., proteins and peptides) based on their isoelectric point (pI). With the advancement in...
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Protocol
Antibodies and Intrabodies Against Huntingtin: Production and Screening of Monoclonals and Single-Chain Recombinant Forms
Antibodies can be extremely useful tools for the field of triplet repeats diseases. These reagents are important for localizing proteins in tissues and they can be used in the isolation and characterization of...
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Protocol
Analysis of Genetic Interactions on a Genome-Wide Scale in Budding Yeast: Diploid-Based Synthetic Lethality Analysis by Microarray
Comprehensive collections of open reading frame (ORF) deletion mutant strains exist for the budding yeast Saccharomyces cerevisiae. With great prescience, these strains were designed with short molecular bar code...
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Protocol
Endogenous Polymerase Assay for the Analysis of Hepatitis B Virus in Transgenic Mice
Hepatitis B virus (HBV) is a human pathogen that can cause severe liver diseases. This virus has a high liver tropism and is often found in large quantities in the sera of patients with HBV. HBV is enveloped a...
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Protocol
Phosphorylation Analysis of Hepatitis B Virus Core Protein in Mammalian Cells
The hepatitis B virus (HBV) C gene encodes the core protein and the precore protein. The core protein, also known as the core antigen, is an important serologic marker for HBV infection. Although this protein ...
