BAC Library Resources for Map-based Cloning and Physical Map Construction in Barley (Hordeum Vulgare L.)

Overview

Journal BMC Genomics

Publisher Biomed Central

Specialty Genetics

Date 2011 May 21

PMID 21595870

Citations 29

Authors

Daniela Schulte

Ruvini Ariyadasa

Bujun Shi

Delphine Fleury

Chris Saski

Michael Atkins

Pieter deJong

Cheng-Cang Wu

Andreas Graner

Peter Langridge

Nils Stein

Affiliations

Soon will be listed here.

Abstract

Background: Although second generation sequencing (2GS) technologies allow re-sequencing of previously gold-standard-sequenced genomes, whole genome shotgun sequencing and de novo assembly of large and complex eukaryotic genomes is still difficult. Availability of a genome-wide physical map is therefore still a prerequisite for whole genome sequencing for genomes like barley. To start such an endeavor, large insert genomic libraries, i.e. Bacterial Artificial Chromosome (BAC) libraries, which are unbiased and representing deep haploid genome coverage, need to be ready in place.

Result: Five new BAC libraries were constructed for barley (Hordeum vulgare L.) cultivar Morex. These libraries were constructed in different cloning sites (HindIII, EcoRI, MboI and BstXI) of the respective vectors. In order to enhance unbiased genome representation and to minimize the number of gaps between BAC contigs, which are often due to uneven distribution of restriction sites, a mechanically sheared library was also generated. The new BAC libraries were fully characterized in depth by scrutinizing the major quality parameters such as average insert size, degree of contamination (plate wide, neighboring, and chloroplast), empty wells and off-scale clones (clones with <30 or >250 fragments). Additionally a set of gene-based probes were hybridized to high density BAC filters and showed that genome coverage of each library is between 2.4 and 6.6 X.

Conclusion: BAC libraries representing >20 haploid genomes are available as a new resource to the barley research community. Systematic utilization of these libraries in high-throughput BAC fingerprinting should allow developing a genome-wide physical map for the barley genome, which will be instrumental for map-based gene isolation and genome sequencing.

Citing Articles

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Leaf Variegation and Impaired Chloroplast Development Caused by a Truncated CCT Domain Gene in Barley.

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