Cytokine‐producing cells are at the center of the adaptive immune responses, and quantifying these cells is an important aspect to build understanding of the immune response.
Rapid progress is being made to understand the regulatory mechanisms that underlie the epigenetic control of gene expression through histone modification. It is now recognized that this plays a major role in normal development and disease.
Mixtures of ascorbate and copper used in certain click chemistry experimental conditions act as oxidizing agents, catalyzing the formation of reactive oxygen species through Fenton and related reactions.
A flow cytometer is made up of many different subsystems that work together to measure the optical properties of individual cells within a sample.
The development of confocal microscopy techniques introduced the ability to optically section fluorescent samples in the axial dimension, perpendicular to the image plane.
Time‐lapse imaging is a rich data source offering potential kinetic information of cellular activity and behavior.
Genome‐wide association studies (GWAS) and sequencing studies are routinely conducted for the identification of genetic variants that are associated with complex traits. Many genetic studies for association mapping include related individuals.
Numerical and structural chromosome abnormalities can be accurately detected in cells from archived tissues using fluorescence in situ hybridization (FISH). This unit describes two common approaches to performing FISH in formalin‐fixed, paraffin‐embedded tissue.
Genomic imbalances involving the telomeric regions of human chromosomes, which contain the highest gene concentration in the genome, are proposed to have severe phenotypic consequences.
Noninvasive prenatal testing (NIPT) represents an outstanding example of how novel scientific discoveries can be quickly and successfully developed into hugely impactful clinical diagnostic tests.
Stranded whole transcriptome RNA‐Seq described in this unit captures quantitative expression data for all types of RNA including, but not limited to, miRNA (microRNA), piRNA (Piwi‐interacting RNA), snoRNA (small nucleolar RNA), lincRNA (large non‐coding intergenic RNA), SRP RNA (signal reco.
Mucopolysaccharidosis type I (MPS I) is a lysosomal storage disorder due to deficiency of alpha iduronidase (IDUA).
Visual analysis is required to perform many biological experiments, from counting colonies to measuring the size or fluorescence intensity of individual cells or organisms.
Proximity ligation assays are a group of protein detection techniques in which reagents with affinity for target proteins, typically antibodies, are coupled to short strands of DNA.
This unit describes Assay for Transposase‐Accessible Chromatin with high‐throughput sequencing (ATAC‐seq), a method for mapping chromatin accessibility genome‐wide.
In order to understand how chromatin complexes function in the nucleus, it is important to obtain a comprehensive picture of their protein, DNA, and RNA components, as well as their mutual interactions.
RNAi has become an essential tool in C. elegans research. This unit describes procedures for RNAi in C.
Advances in modern optical microscopy have provided unparalleled tools to study intracellular structure and function, yet visualizing lipid molecules within a cell remains challenging.
This unit describes methods for non‐isotopic RNA in situ hybridization on embryonic mouse sections. These methods can be used to follow the spatiotemporal dynamics of gene expression in an embryonic tissue of interest. They involve the use of labeled (e.g.
A better understanding of mechanisms governing receptor insertion to the plasma membrane (PM) requires an experimental approach with excellent spatial and temporal resolutions.
The striatum mediates a variety of functions including movement, decision‐making, motivation, and reward learning. In vivo recording is a powerful technique that allows for the interrogation of these striatal functions while an animal is awake and behaving.
Fast‐scan cyclic voltammetry (FSCV) is an electrochemical technique that permits the in vivo measurement of extracellular fluctuations in multiple chemical species.
This protocol details a free‐operant avoidance paradigm that has been developed to evaluate the relative contribution of different sources of reinforcement of avoidance behavior that may play an important role in the development and maintenance of human anxiety disorders.