遗传学实验技术

The Long serial analysis of gene expression (SAGE) protocol generates ditags from tags with overlapping overhangs, thereby increasing the probability of duplicate ditag formation in LongSAGE. In this chapter, a tool is presented that facilitates the analysis of duplicate ditags in Lo ...

Single-nucleotide polymorphism (SNP) mapping is the easiest and most reliable way to map genes in Caenorhabditis elegans. SNPs are extremely dense and usually have no associated phenotype, making them ideal markers for mapping. SNP mapping has three steps. First, recombinant mutant an ...

The methods used by the Caenorhabditis elegans Gene Knockout Consortium are conceptually simple. One does a chemical mutagenesis of wild-type C. elegans, and then screens the progeny of the mutagenized animals, in small mixed groups, using polymerase chain reaction (PCR) to identify pop ...

The COPAS™ Biosorter is a flow cytometer designed to accommodate large objects the size of Caenorhabditis elegans. This instrumentation brings high-speed automated analysis and sorting to this small model organism. The Biosort system optically analyzes and sorts living multicel ...

Automated systems for recording and analyzing behavior have many applications for the study of neurobiology in Caenorhabditis elegans. In particular, machine-based approaches allow for precise quantitative definitions of behavioral phenotypes that have traditionally ...

The nematode Caenorhabditis elegans is an extraordinarily powerful model organism for the application of functional genomic approaches. Two such approaches, whole genome microarray analysis and genome-wide RNA interference (RNAi)-mediated phenotypic screening, are hi ...

This chapter describes four methods for delivery of double-stranded RNA to Caenorhabditis elegans (injection, feeding, soaking, and in vivo delivery), and suggests schemes that should facilitate detection of specific gene silencing.

Double-stranded RNA (dsRNA)-induced gene silencing in Caenorhabditis elegans involves the manufacture and delivery of defined sequences of dsRNA to the organism, followed by a careful monitoring for loss-of-function phenocopies in treated animals. In this chapter, we describe h ...

The establishment of Caenorhabditis elegans as a “model organism” began with the efforts of Sydney Brenner in the early 1960s. Brenner’s focus was to find a suitable animal model in which the tools of genetic analysis could be used to define molecular mechanisms of development and nervous system ...

One major postgenomic challenge is to characterize the epigenomes that control genome functions. The epigenomes are mainly defined by the specific association of nonhistone proteins with chromatin and the covalent modifications of chromatin, including DNA methylation and pos ...

Combining serial analysis of gene expression (SAGE) with pyrophosphatase-based ultra-high-throughput DNA sequencing provides increased sensitivity and cost-effective gene expression profiling. The combined techniques obviate the formation and cloning of concate ...

Serial analysis of gene expression (SAGE) requires the sequencing of DNA. The principal cost of SAGE is largely determined by the cost of sequencing. Therefore, it is important to have access to a robust and affordable sequencing system. Here, we describe such a system based on the sequencing of ampl ...

As a tool for high-throughput, quantitative gene expression analysis, serial analysis of gene expression (SAGE) is one of the most powerful techniques. However, the short size of tags (14bp) has hindered the application of SAGE to a vast majority of eukaryotes without sufficient genomic res ...

In order to generate serial analysis of gene expression (SAGE) libraries from very small samples such as microdissected cells, the starting material must first be amplified via PCR or linear amplification of RNA. In microarray experiments, it has been shown that linear amplification of RNA c ...

Serial analysis of gene expression (SAGE) is a powerful technique for large-scale transcriptome analysis in eukaryotes. However, technical difficulties in the SAGE library construction, such as low concatemer cloning efficiency, small concatemer size, and a high level of empty clo ...

Serial analysis of gene expression (SAGE) is a high-throughput method for global gene expression analysis that allows the quantitative and simultaneous analysis of a large number of transcripts. SAGE is a digital method and its sensitivity depends only on the number of tags sequenced. Furt ...

Data analysis of serial analysis of gene expression (SAGE) tag experiments begins with the extraction of tags from single-pass sequence files of ditag concatemers. When using DNA base quality values generated during base calling, it is possible to control the false-positive discovery r ...

Many newly identified gene products from completely sequenced genomes are difficult to characterize in the absence of sequence homology to known proteins. In such a scenario, the context of the proteins’ functional associations can be used for annotation; overrepresented function ...

The combinatorial control of gene regulatory switches involves both transcription factor (TF) complexes and associated epigenetic modifications to the chromatin template. The novel highthroughput technologies, such as Chromatin ImmunoPrecipitation ChIP-chip, have ...

Detailed instruction is described for mapping unstructured, free text data into common biomedical concepts (drugs, diseases, anatomy, and so on) found in the Unified Medical Language System using MetaMap Transfer (MMTx). MMTx can be used in applications including mining and inferring ...