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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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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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Rapid Whole Genome Sequencing for Diagnosis of Single Locus Genetic Diseases in Critically Ill Children
Upon admission to intensive care units (ICU), the differential diagnosis of almost all infants with diseases of unclear etiology includes single...
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Flow Cytometry-Based Detection of Siglec Ligands
Siglecs are a family of immunomodulatory cell surface receptors present on white blood cells. Binding to cell surface sialic acid-containing glycans...
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Separation and Visualization of Glycans by Fluorophore-Assisted Carbohydrate Electrophoresis
Fluorophore-assisted carbohydrate electrophoresis (FACE) is a method in which a fluorophore is covalently attached to the reducing end of...
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Isolation and Preparation of Extracellular Proteins from Lignocellulose-Degrading Fungi for Comparative Proteomic Studies Using Mass Spectrometry
Fungi utilize a unique mechanism of nutrient acquisition involving extracellular digestion. To understand the biology of these microbes, it is...
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Generation of Amber Suppression Cell Lines Using CRISPR-Cas9
Genetic code expansion via amber suppression allows cotranslational, site-specific introduction of nonnatural chemical groups into proteins in the...
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Cell Subset-Specific Chimerism Testing by Short Tandem Repeats Analysis for Engraftment Monitoring After Hematopoietic Stem Cell Transplantation
Chimerism is the unique state when cells from genetically different individuals coexist. Chimerism testing allows measuring the donor and recipient...
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MYD88 L265P Mutation Detection by ddPCR: Recommendations for Screening and Minimal Residual Disease Monitoring
MYD88L265P is a gain-of-function mutation, arising from the missense alteration c.794T>C, that frequently occurs in B-cell malignancies such as...
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Genetically Encoded 1,2-Aminothiol for Site-Specific Modification of a Cellular Membrane Protein via TAMM Condensation
Site-specific modification of proteins has wide applications in probing and perturbing biological systems. A popular means to achieve such a...
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Capture Methylation-Sensitive Restriction Enzyme Sequencing (Capture MRE-Seq) for Methylation Analysis of Highly Degraded DNA Samples
Understanding the impact of DNA methylation within different disease contexts often requires accurate assessment of these modifications in a...
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Long-Read Metagenomics and CAZyme Discovery
Microorganisms play a primary role in regulating biogeochemical cycles and are a valuable source of enzymes that have biotechnological applications,...
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EOSAL–CNV for Easy and Rapid Detection of CNVs by Fragment Analysis
Copy number variations (CNVs) are a type of genetic variation involving from 50 base pairs (bps) to millions of bps and, in a general point of view,...
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Rapid Cycle and Extreme Polymerase Chain Reaction
Rapid cycle polymerase chain reaction (PCR) amplifies DNA in 10–30 min, while extreme PCR is complete in less than 1 min. These methods do not...
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Isothermal Titration Calorimetry for Quantification of Protein–Carbohydrate Interactions
Isothermal titration calorimetry allows the determination of thermodynamic parameters for the interaction between a protein and mono- or...
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Biomechanical Weakening of Paper and Plant Cell Walls by Bacterial Expansins
Expansins are proteins that loosen plant cell walls but lack enzymatic activity. Here we describe two protocols tailored to measure the biomechanical...
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Identification of Genes Involved in the Degradation of Lignocellulose Using Comparative Transcriptomics
Lignocellulosic biomass represents an abundant, renewable resource that can be used to produce biofuels, low-cost livestock feed, and high-value...
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Reprogramming Initiator and Nonsense Codons to Simultaneously Install Three Distinct Noncanonical Amino Acids into Proteins in E. coli
Multiple noncanonical amino acids can be installed into proteins in E. coli using mutually orthogonal aminoacyl-tRNA synthetase and tRNA pairs. Here...
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Small-Molecule Phosphine Activation of Protein Function in Zebrafish Embryos with an Expanded Genetic Code
Conditional control of protein function in a living model organism is an important tool for studying the effects of that protein during development...
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Managing a PCR Contamination Event in a Molecular Pathology Laboratory
Contamination in a molecular laboratory may lead to erroneous results with potential to cause patient harm if not promptly identified and corrected....