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CHORI-201 Maize B73 BAC Library

The CHORI-201 Maize BAC Library has been constructed in our laboratory by Qing Dan Chen, Chung Li Shu and Kazutoyo Osoegawa. The library is being used for physical mapping and sequencing of the maize genome. In genomic databases, the library has (unfortunately) been re-named to" ZMMBBc" (see for instance the Arizona Genomics Institute's web site). In databases, the BACs can be found with a prefix "c" preceding microtiter dish number, row and column intersection. For instance, "c0460G23" is a particular clone in the database. When ordering BACs, the clone name will need to be translated into the NCBI format: http://www.ncbi.nlm.nih.gov/genome/clone/nomen.html. Hence, BAC "c0460G23" will become "CH201-460G23" in our version. 

Library construction details: Genomic DNA was isolated from B73 nuclei. The first segment was constructed using DNA partially digested with a combination of EcoRI and EcoRI Methylase, the second segment was constructed using MboI partially digested DNA. Size selected DNA was cloned into the pTARBAC2.1 vector between the EcoRI sites and into the pTARBAC1.3 vector between the BamHI sites. The ligation products were transformed into DH10B electrocompetent cells (BRL Life Technologies). BAC clones have been arrayed into 288 384-well microtiter dishes for each library segment and gridded onto 6 22x22cm nylon high-density filters for each library segment for screening by probe hybridization. Each hybridization membrane represents over 18,000 distinct maize BAC clones, stamped in duplicate. Library construction was supported by sub-contracts from University of Missouri for segment 1 and Rutgers University for segment 2, through NSF grants awarded to Jack Gardner and Joachim Messing, respectively.

Provisonal data for CHORI-201 Maize B73 BAC library:

Data on the CHORI-201 clone average insert size  of Segment 1 and Segment 2 have been determined by Pulsed Field Gel Electrophoresis. Clone size distribution for each segment has been plotted graphically. While analyzing clones using pulse-field electrophoresis to determine the average insert size, non-insert clones containing a small deleted vector fragment consistent with sucrose resistance were observed. Further in depth characterization of the library is on going in our lab and data will be updated on our web page periodically.

Please direct questions concerning this library to either Pieter J. de Jong or Kazutoyo Osoegawa.

The library is available in several formats. Individual clones, and high-density hybridization filters  are obtainable. For ordering and shipping details, please view the ordering and pricing information  page.

Academic and commercial users interested in a copy of the BAC library should contact Pieter J. de Jong ( pdejong@chori.org ), fax: (510) 450-7924).

Yim, Y. S., Davis , G. L., Duru, N. A., Musket, T. A., Linton, E. W., Messing, J. W., McMullen, M. D., Soderlund, C. A., Polacco, M. L., Gardiner, J. M., Coe, E. H., Jr. (2002) Characterization of three maize bacterial artificial chromosome libraries toward anchoring of the physical map to the genetic map using high-density bacterial artificial chromosome filter hybridization. Plant Physiol 130, 1686-1696.