Detecting genes
著者
書誌事項
Detecting genes
（Genome analysis : a laboratory manual / editors, Eric D. Green ... [et al.], v. 2）
Cold Spring Harbor Laboratory Press, c1998
- : comb
- : cloth
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注記
Includes bibliographical references and index
内容説明・目次
内容説明
Assuming only a basic knowledge of molecular biology, this is the second in a series of manuals which explains how to clone, manipulate, analyze and sequence large segments of DNA, and relate expressed sequence to phenotypic variation. The techniques are written for application to animal DNA as well as human genomes. They deal plainly with sources of failure, and solutions.
目次
- Strategies for gene discovery in mammalian systems
- gene identification
- mapping strategies
- isolation of transcribed sequences
- proving a candidate gene is casually mutated
- future developments. Constructing and screening normalized cDNA libraries: primary uses and applicatons for cDNA libraries
- overview of the methods for synthesizing double-stranded cDNA
- cloning vectors for construction of cDNA libraries
- overview of the procedure for constructing directionally cloned cDNA libraries
- problems commonly encountered in cDNA libraries
- cloning efficiencies required to ensure that a representative cDNA library is obtained
- theoretical considerations in normalizing cDNA populations
- overview of the methods for constructing normalized cDNA libraries
- methods for isolating RNA - protocols
- construction of oligo (dt)-pimed, directionally clone cDNA libraries in phagemid vectors - protocols
- normalization of directionally cloned cDNA libraries - protocols
- screening normalized cDNA libraries constructed in phagemid vectors. Direct cDNA. Selection: overview of direct cDNA selection
- choosing sources of cloned genomic DNA and cDNA for direct cDNA selection
- preparation of biotinylated, clone genomic DNA - protocols
- preparation of cDNA - protocols
- hybridization reactions and enrichment of the cDNA - protocols
- confirming that cDNA products arise from transcriptionally active genes in the genomic region of interest. Exon trapping: strategies for trapping intact exons
- protocols for trapping intact exons
- internal exon trapping using pSPL3 - protocols
- 3-terminal exon trapping using pTAG4 - protocols
- analysis and identification of trapped exons. Gene detection by the identification of cpG islands: comparison of the cpGislands and vertebrate genomic DNA
- cpG islands as markers for genes
- evolution of approaches for identifying CpG islands
- recently developed techniques for identifying CpG islands
- advantages of isolating CpG islands
- advantages of isolating CpG islands as an approach to gene identification
- choosing a strategy for isolating genes associated with CpG islands
- combining approaches for gene isolation
- using RCREs to identify CpG-rich sequences in cloned genomic DNA - protocols
- data analysis for identification of genes associated with CpG islands from cloned genomic DNA - protocols
- using a column that binds methylated DNA to purify CpG islands from uncloned genomics DNA - protocols. Detection of DNA variation: overview of DNA variation
- methods for detecting DNA variation
- choosing a suitable approach for mutation detection
- the SSCP method
- protocols for detecting DNA variation using SSCP analysis
- the DGGE method
- equipment and protocols for detecting DNA variation using DGGE: RNase cleavage at mismatches
- protocols for detecting DNA variation usng Rnase cleavage
- the CCM method
- protocols for detecting DNA variation using CCM
- direct sequencing of uncloned PCR products. (Part contents)
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