遗传学实验技术

Next generation sequencing technologies may now be applied to the study of transcriptomics. RNA-Seq or RNA sequencing employs high-throughput sequencing of complementary DNA fragments delivering a transcriptional profile. In this chapter, we aim to provide a starting point for Xe ...

Gene expression data for Xenopus are collected and curated in diverse forms and locations. The intention of this chapter is to give the reader a guide to the publicly accessible databases where these data can be found and an idea of the current scope and limitations of the data in these resources. Instru ...

Alternative splicing, the process by which distinct mature mRNAs can be produced from a single primary transcript, is a key mechanism to increase the organism complexity. The generation of alternative splicing pattern is a means to expand the proteome diversity and also to control gene expr ...

The immunoaffinity isolation of protein complexes is an essential technique for the purification and �concentration of protein complexes from cells and tissues. In this chapter we present the methodologies for the purification of proteins and protein complexes from Xenopus laev ...

Proteomic characterization of protein complexes leverages the versatile platform of liquid chromatography-tandem mass spectrometry to elucidate molecular and cellular signaling processes underlying the dynamic regulation of macromolecular assemblies. Here, we d ...

In situ hybridization involves the hybridization of an antisense RNA probe to an mRNA transcript and it is a powerful method for the characterization of gene expression in tissues, organs, or whole organisms. Performed as a whole mount (WISH), it allows the detection of mRNA transcripts in three d ...

Antibody-based detection of protein distribution patterns both in wholemount and on sections revolutionized Xenopus research and ushered in the visual-based era of Xenopus data presentation. The ability to view the distribution of a gene product throughout an embryo makes it possi ...

Fluorescence in situ hybridization (FISH) is a commonly used technique for the visualization of whole chromosomes or subchromosomal regions, such as chromosome arms, bands, centromeres, or single gene loci. FISH is routinely performed on chromosome spreads, as well as on three-dimens ...

Insight into the normal and abnormal function of an interphase nucleus can be revealed by using fluorescence in situ hybridization (FISH) to determine chromosome copy number and/or the nuclear position of loci or chromosome territories. FISH has been used extensively in studies of mouse a ...

This chapter concentrates on the use of fluorescence in situ hybridization (FISH) for genomic investigations in the laboratory rat (Rattus norvegicus). The selection of protocols included in the chapter has been inspired by a comprehensive range of previously published molecular c ...

A common problem in research laboratories that study the mammalian embryo is the limited supply of live material. For this reason, new methods are constantly being developed and existing methods for in vitro models using cells in culture are being adapted to represent embryogenesis. Three- ...

The organization of eukaryotic chromosomes into euchromatin and heterochromatin represents an enigmatic aspect of genome evolution. Constitutive heterochromatin is a basic, yet still poorly understood component of eukaryotic genomes and its molecular characterizati ...

The application of fluorescence in situ hybridization (FISH) for the mapping of single copy genes onto homologous chromosome has been integral to vast number genome sequencing projects, such as that of mouse and human. The chromosomes of these organisms are well-studied and are the staple re ...

Fluorescence in situ hybridization (FISH) provides an effective means to delineate chromosomes and their subregions during all stages of the cell cycle. This makes FISH particularly useful for studying chromosome behavior in species with minute genomes and/or poor chromosome con ...

Fluorescence in situ hybridization with rRNA targeted oligonucleotide probes is nowadays one of the core techniques in microbial ecology, allowing the identification and quantification of microbial cells in environmental samples in situ. Next to the classic FISH protocol, which ...

The field of virology is undergoing a revolution as diagnostic tests and new therapies are allowing clinicians to treat, monitor, and predict outcomes of viral diseases. The majority of these techniques, however, destroy the factory of viral production and the information inherent in the r ...

Genomes are spatially highly organized within interphase nuclei. Spatial genome organization is increasingly linked to genome function. Fluorescence in situ hybridization (FISH) allows the visualization of specific regions of the genome for spatial mapping. While most gene lo ...

The detection of genetic abnormalities in paraffin sections by fluorescence in situ hybridization (FISH) is widely used in clinical practice to detect amplification of the ERB2 gene in breast carcinoma and various chromosomal translocations in lymphomas and soft tissue tumors. How ...

The frog Xenopus laevis has for more than 60 years served as a model system for the study of vertebrate embryogenesis, molecular and cell biology, and physiology. Recently, there has been great interest in the related species Xenopus tropicalis, in part because it is diploid, unlike the allotetra ...

A spectacular advantage of Xenopus tropicalis is the ease with which diploid embryos can be generated year round. By the simple administration of human chorionic gonadotropin, an investigator can generate many hundreds of synchronized embryos by in vitro fertilization or thousands of ...