Gene Isolation And Mapping Protocols (methods In Molecular Biology)
by Jacqueline Boultwood /
1996 / English / PDF
20.4 MB Download
Over the past 10 years great progress has been made in the
development of efficient techniques for both gene isolation and
mapping. The identifica tion and isolation of transcribed
sequences from large chromosomal regions are central to the human
genome mapping project. Techniques for isolating novel cDNAs have
applications both in the overall construction and integra tion of
long-range physical and transcription maps and in the
identification of disease genes. A number of different techniques
for the isolation of cDNAs from mam malian genomes have been
developed, including screening "zoo" blots, the use of large
genomic clones (YACs or cosmids) for hybridization against cDNA
libraries, and CpG island mapping. More recently two highly
efficient tech niques have been introduced: exon trapping, based
on the presence of exon splice sites, and direct selection, based
on the enrichment of selected cDNAs using immobilized YACs or
cosmids. Leading researchers in the field have contributed chapters
detailing the practical procedures for these and other widely used
methods. The most rapid progress presently being made in the field
of gene isolation concerns the partial sequencing of cDNA clones
from one or both ends to produce expressed sequence tags (ESTs).
Indeed, by Octo ber 1995, the EST division of Genbank (dbEST)
contained a total of approxi mately 270,000 human EST sequences
accounting for almost half the number of sequence entries in
Genbank.
Over the past 10 years great progress has been made in the
development of efficient techniques for both gene isolation and
mapping. The identifica tion and isolation of transcribed
sequences from large chromosomal regions are central to the human
genome mapping project. Techniques for isolating novel cDNAs have
applications both in the overall construction and integra tion of
long-range physical and transcription maps and in the
identification of disease genes. A number of different techniques
for the isolation of cDNAs from mam malian genomes have been
developed, including screening "zoo" blots, the use of large
genomic clones (YACs or cosmids) for hybridization against cDNA
libraries, and CpG island mapping. More recently two highly
efficient tech niques have been introduced: exon trapping, based
on the presence of exon splice sites, and direct selection, based
on the enrichment of selected cDNAs using immobilized YACs or
cosmids. Leading researchers in the field have contributed chapters
detailing the practical procedures for these and other widely used
methods. The most rapid progress presently being made in the field
of gene isolation concerns the partial sequencing of cDNA clones
from one or both ends to produce expressed sequence tags (ESTs).
Indeed, by Octo ber 1995, the EST division of Genbank (dbEST)
contained a total of approxi mately 270,000 human EST sequences
accounting for almost half the number of sequence entries in
Genbank.
