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Nucleic Acids Research, 2000, Vol. 28, No. 24 e106
© 2000 Oxford University Press

Non-gridded library: a new approach for BAC (bacterial artificial chromosome) exploitation in hexaploid wheat (Triticum aestivum)

Zhiying Ma1,2, Song Weining3, Peter J. Sharp4 and Chunji Liu1,*

1CSIRO Plant Industry, 306 Carmody Road, St Lucia, Queensland 4067, Australia, 2Faculty of Agriculture, Hebei Agricultural University, Baoding, Hebei, People’s Republic of China, 3Leslie Research Centre, 13 Holberton Steet, Toowoomba, Queensland 4350, Australia and 4Plant Breeding Institute, Cobbitty, NSW 2570, Australia

The feasibility of exploiting non-gridded bacterial artificial chromosome (BAC) libraries and some major factors affecting the efficiency of handling such libraries were studied in hexaploid wheat. Even for a bacterial culture containing only 55% recombinants, some 2000 BAC clones with inserts ranging from 45 to 245 kb could be pooled. The pooled BAC clones could be amplified by culturing for up to 6 h without losing any target clones. These results imply that even for hexaploid wheat, which has an extremely large genome, some 250 pools are sufficient for a BAC library that should satisfy many research objectives. This non-gridded strategy would dramatically reduce the cost and make robotic equipment non-essential in exploiting BAC technology. To construct a representative library and to minimise clone competition, thawing and re-freezing ligation mixtures and bacterial cultures should be avoided in BAC library construction and application.

* To whom correspondence should be addressed. Tel: +61 7 3214 2223; Fax: +61 7 3214 2848; Email: chunji.liu{at}pi.csiro.au


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I. Ratnayaka, M. Baga, D. B. Fowler, and R. N. Chibbar
Construction and Characterization of a BAC Library of a Cold-Tolerant Hexaploid Wheat Cultivar
Crop Sci., June 24, 2005; 45(4): 1571 - 1577.
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