GrainGenes Reference Report: GNM-42-1176
Reference
GNM-42-1176
Title
Construction and characterization of a bacterial artificial chromosome (BAC) library for the A genome of wheat
Journal
Genome
Year
1999
Volume
42
Pages
1176-1182
Author
Lijavetzky D
[ Show all 6 ]
Abstract
Summary: A genomic bacterial artificial chromosome (BAC) library of the A genome of wheat has been constructed. Triticum monococcum accession DV92 was selected for this purpose because it is a cultivated diploid wheat and one of the parental lines used in the construction of a saturated genetic map. Leaves from this accession were used to isolate high-molecular-weight DNA from nuclei. This DNA was partially digested with restriction enzyme Hind III, subjected to double size selection, electroeluted and cloned into the pINDIGO451 BAC vector. The library consists of 276 480 clones with an average insert size of 115 kb. Excluding the 1.33% of empty clones and 0.14% of clones with chloroplast DNA, the coverage of this library is 5.6 genome equivalents. With this genome coverage the probability of having any DNA sequence represented in this library is higher than 99.6%. Clones were sorted in 720 384-well plates and blotted onto 15 high-density filters. High-density filters were screened with several single or low-copy clones and five positive BAC clones were selected for further analysis. Since most of the T. monococcum BAC ends included repetitive sequences, a modification was introduced into the classical end-isolation procedure to select low copy sequences for chromosome walking
External Databases
Pubmed: 10659785
Keyword
GrainGenes is a product of the Agricultural Research Service of the US Department of Agriculture.
GrainGenes Reference Report: GNM-42-1176
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GrainGenes is a product of the Agricultural Research Service of the US Department of Agriculture. | |||