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SARS-CoV-2 Sequencing for Variant Surveillance
In this chapter, next-generation sequencing of the entire viral genome of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is described....
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Immune Cell Lineage-Specific Chimerism Testing by Next-Generation Sequencing for Engraftment Monitoring After Allogeneic Hematopoietic Stem Cell Transplantation
Chimerism is an unusual state in which a person’s body comprises cells from genetically different people. Chimerism testing allows monitoring for the...
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Encoding Noncanonical Amino Acids into Phage Displayed Proteins
Phage display facilitates the evolution of peptides and proteins for affinity selection against targets, but it is mostly limited to the chemical...
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Tuberculosis Diagnosis Using Isothermal Nucleic Acid Amplification in a Paper-and-Plastic Device
Nucleic acid (DNA/RNA) amplification technologies are indispensable for applications like disease diagnostics, forensics, epidemiology, evolutionary...
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Ultrasensitive Reverse Transcriptase Loop-Mediated Isothermal Amplification (US-LAMP)-Based Detection of Malaria Infection from Dried Blood Spots
Submicroscopic malaria diagnosis requires highly sensitive tools instead of the conventional microscopy and rapid diagnostic tests (RDTs). While...
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Quantifying CBM–Carbohydrate Interactions Using Microscale Thermophoresis
Microscale thermophoresis (MST) is an emerging technology for studying a broad range of biomolecular interactions with a high sensitivity. The...
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Site-Specific Incorporation of Sulfotyrosine into Proteins in Mammalian Cells
Protein tyrosine O-sulfationposttranslational modifications (PTMs)tyrosine O-sulfation (PTS) plays a crucial role in numerous extracellular...
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Genetically Encoded Noncanonical Amino Acids in Proteins to Investigate Lysine Benzoylation
Posttranslational modifications (PTMs) of lysine residues are major regulators of gene expression, protein–protein interactions, and protein...
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Protein Expression with Biosynthesized Noncanonical Amino Acids
Natural proteins are normally made by 20 canonical amino acids. Genetic code expansion (GCE) enables incorporation of diverse chemically synthesized...
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A Brief Practical Guide to PCR
Polymerase chain reaction (PCR) has been a powerful molecular biology tool since the mid-1980s. Millions of copies of specific sequence regions of...
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Loop-Mediated Isothermal Amplification on Paper Microfluidic Chips for Highly Sensitive and Specific Zika Virus Detection Using Smartphone
Zika virus (ZIKV) infection may cause serious birth defects and is a critical concern for women of child-bearing age in affected regions. A simple,...
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Measuring Enzyme Kinetics of Glycoside Hydrolases Using the 3,5-Dinitrosalicylic Acid Assay
Use of the 3,5-dinitrosalicylic acid reagent allows the simple, rapid quantification of reducing sugars. The method can be used for analysis of...
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Nuclease Enrichment and qPCR Detection of Rare Nucleotide Variants
The emergence of circulating DNA analysis in blood during the past decade has responded to the need for noninvasive alternatives to classical tissue...
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CBMs as Probes to Explore Plant Cell Wall Heterogeneity Using Immunocytochemistry
Immunocytochemistry is a widely used technique to localize antigen within intact tissues. Plant cell walls are complex matrixes of highly decorated...
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Affinity Electrophoresis for Analysis of Catalytic Module–Carbohydrate Interactions
Affinity electrophoresis has long been used to study the interaction between proteins and large soluble ligands. The technique has been found to have...
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A Rapid Protocol for Preparing 8-Aminopyrene-1,3,6-Trisulfonate-Labeled Glycans for Capillary Electrophoresis-Based Enzyme Assays
Purified glycan standards are required for glycan arrays, characterizing substrate specificities of glycan-active enzymes, and to serve as...
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Creating Selenocysteine-Specific Reporters Using Inteins
The selenium moiety in selenocysteine (Sec) imparts enhanced chemical properties to this amino acid and ultimately the protein in which it is...
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Genetic Encoding of a Fluorescent Noncanonical Amino Acid as a FRET Donor for the Analysis of Deubiquitinase Activities
The genetic code expansion technology enables the genetic encoding of fluorescent noncanonical amino acids (ncAAs) for site-specific fluorescent...
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Next-Generation Sequencing Informatic Architecture Considerations
Clinically relevant sequencing methodologies continue to expand in number, diversity, complexity, and scale. This evolving and varied landscape...