NDLI: Direct targeting and rapid isolation of BAC clones spanning a defined chromosome region

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Direct targeting and rapid isolation of BAC clones spanning a defined chromosome region

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Functional & Integrative Genomics : Volume 5, Issue 1, January 2005

Functional & Integrative Genomics : Volume 4

Functional & Integrative Genomics : Volume 3

Functional & Integrative Genomics : Volume 2

Functional & Integrative Genomics : Volume 1

Direct targeting and rapid isolation of BAC clones spanning a defined chromosome region

Content Provider	Springer Nature Link
Author	Isidore, Edwige Scherrer, Beatrice Bellec, Arnaud Budin, Karine Faivre Rampant, Patricia Waugh, Robbie Keller, Beat Caboche, Michel Feuillet, Catherine Chalhoub, Boulos
Copyright Year	2005
Abstract	To isolate genes of interest in plants, it is essential to construct bacterial artificial chromosome (BAC) libraries from specific genotypes. Construction and organisation of BAC libraries is laborious and costly, especially from organisms with large and complex genomes. In the present study, we developed the pooled BAC library strategy that allows rapid and low cost generation and screening of genomic libraries from any genotype of interest. The BAC library is constructed, directly organised into a few pools and screened for BAC clones of interest using PCR and hybridisation steps, without requiring organization into individual clones. As a proof of concept, a pooled BAC library of approximately 177,000 recombinant clones has been constructed from the barley cultivar Cebada Capa that carries the Rph7 leaf rust resistance gene. The library has an average insert size of 140 kb, a coverage of six barley genome equivalents and is organised in 138 pools of about 1,300 clones each. We rapidly established a single contig of six BAC clones spanning 230 kb at the Rph7 locus on chromosome 3HS. The described low-cost cloning strategy is fast and will greatly facilitate direct targeting of genes and large-scale intra- and inter-species comparative genome analysis.
Starting Page	97
Ending Page	103
Page Count	7
File Format	PDF
ISSN	1438793X
Journal	Functional & Integrative Genomics
Volume Number	5
Issue Number	2
e-ISSN	14387948
Language	English
Publisher	Springer-Verlag
Publisher Date	2005-01-22
Publisher Place	Berlin, Heidelberg
Access Restriction	One Nation One Subscription (ONOS)
Subject Keyword	Barley Pooled BAC library Screening Targeting genes of interest
Content Type	Text
Resource Type	Article
Subject	Genetics Medicine

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1	Ministry of Education (GoI), Department of Higher Education	Sanctioning Authority	https://www.education.gov.in/ict-initiatives
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3	National Digital Library of India Office, Indian Institute of Technology Kharagpur	The administrative and infrastructural headquarters of the project	Dr. B. Sutradhar bsutra@ndl.gov.in
4	Project PI / Joint PI	Principal Investigator and Joint Principal Investigators of the project	Dr. B. Sutradhar bsutra@ndl.gov.in Prof. Saswat Chakrabarti will be added soon
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