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United States Department of Agriculture

Agricultural Research Service

Title: Construction and Characterization of a Bacterial Artificial Chromosome Library from Soybean

item Marek, Laura - ISU
item Shoemaker, Randy

Submitted to: Meeting Abstract
Publication Type: Abstract Only
Publication Acceptance Date: August 30, 1996
Publication Date: N/A

Technical Abstract: A bacterial artificial chromosome (BAC) library suitable for map-based cloning and physical mapping has been constructed in soybean. The BAC vector, capable of maintaining large insert DNA, was developed by Shizuya et al. (P.N.A.S. 89:8794 1992) for use in human genome mapping. The first plant BAC work was in Rod Wing's laboratory at Texas A&M (Woo et al. NAR 22:4922 1994). Our soybean BAC library uses the pBeloBAC 11 vector. Large insert DNA was isolated from Hind III partial digests of megabase DNA prepared from nuclei embedded in agarose microbeads. The nuclei were prepared from young leaves of the Williams 82 soybean cultivar. A random selection of 160 BACs generated from two ligations prepared from DNA size-selected from partial digests of two different microbead preparations ranged in size from 40 to 350 kb with an average insert size of 150 kb. The BAC library is stored as individually picked clones representing 4-5 genome equivalents (approximately 40,000) in 384 well microtiter dishes. We have prepared pooled DNA samples suitable for screening the library with PCR based technologies as well as membranes for screening by hybridization methods.

Last Modified: 4/22/2015
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