Tags

Type your tag names separated by a space and hit enter

Construction and characterization of a bacterial artificial chromosome (BAC) library for the A genome of wheat.
Genome. 1999 Dec; 42(6):1176-82.G

Abstract

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.

Authors+Show Affiliations

Department of Agronomy & Range Science, University of California, Davis 95616-8515, USA.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.

Language

eng

PubMed ID

10659785

Citation

Lijavetzky, D, et al. "Construction and Characterization of a Bacterial Artificial Chromosome (BAC) Library for the a Genome of Wheat." Genome, vol. 42, no. 6, 1999, pp. 1176-82.
Lijavetzky D, Muzzi G, Wicker T, et al. Construction and characterization of a bacterial artificial chromosome (BAC) library for the A genome of wheat. Genome. 1999;42(6):1176-82.
Lijavetzky, D., Muzzi, G., Wicker, T., Keller, B., Wing, R., & Dubcovsky, J. (1999). Construction and characterization of a bacterial artificial chromosome (BAC) library for the A genome of wheat. Genome, 42(6), 1176-82.
Lijavetzky D, et al. Construction and Characterization of a Bacterial Artificial Chromosome (BAC) Library for the a Genome of Wheat. Genome. 1999;42(6):1176-82. PubMed PMID: 10659785.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Construction and characterization of a bacterial artificial chromosome (BAC) library for the A genome of wheat. AU - Lijavetzky,D, AU - Muzzi,G, AU - Wicker,T, AU - Keller,B, AU - Wing,R, AU - Dubcovsky,J, PY - 2000/2/5/pubmed PY - 2000/4/25/medline PY - 2000/2/5/entrez SP - 1176 EP - 82 JF - Genome JO - Genome VL - 42 IS - 6 N2 - 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. SN - 0831-2796 UR - https://www.unboundmedicine.com/medline/citation/10659785/Construction_and_characterization_of_a_bacterial_artificial_chromosome__BAC__library_for_the_A_genome_of_wheat_ DB - PRIME DP - Unbound Medicine ER -