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BAC Library Construction

 Home > Resources > Libraries
  CHORI-242 Porcine BAC Library   
  Background information

The CHORI-242 porcine (Sus scrofa) BAC library has been constructed by Baoli Zhu in Pieter de Jong’s laboratory at BACPAC Resources, Children’s Hospital Oakland Research Institute. The preparation of the library followed the cloning approach developed in our laboratory (Osoegawa et al., 1998). Blood sample was obtained from a single Duroc female (Herd ID 2-14) located at the UIUC Moorman Swine Research farm through Dr. Jonathan E. Beever (Department of Animal Sciences, University of Illinois at Urbana-Champaign). High-molecular-weight DNA was isolated from white blood cells. Agarose embedded high molecular weight DNA was partially digested with MboI and size fractionated DNA was cloned into the pTARBAC1.3 vector between the BamHI sites. The ligation products were transformed into DH10B (T1 resistant) electrocompetent cells (Invitrogen). The library has been arrayed into 384-well microtiter dishes and also gridded onto eleven 22x22cm nylon high-density filters for screening by probe hybridization. Each hybridization membrane represents over 18,000 distinct porcine BAC clones, stamped in duplicate. Library characterization was performed by Qing Cao and Kazutoyo Osoegawa. BAC library construction was funded by a grant from USDA (99-35205-8537).

Provisional data for CHORI-242 Procine BAC Library:

Segment Cloning Vector DNA Restriction enzyme Plate Number Total Plates Empty Wells Empty Wells (%)
1 pTARBAC1.3 White Blood Cell MboI 1-288 288 1,126 1.02
2 pTARBAC1.3 White Blood Cells MboI 289-528 240 952 1.03
Total Library       1-528 528 2,078 1.02


Segment Non-insert clones (%) Non-insert clones Non-Recombinant Clones (pUC) Recombinant clones Insert Size (Average) Redundancy (Genome Size: 3000 Mb)
1 2.08 Approx. 2,300 0 Approx. 107,166 177Kbp 6.3
2 2.08 Apporx. 1,917 0 Approx. 89,291 170Kbp 5.1
Total Library 2.08 Apporx. 4,217 0 Approx. 196,457 173Kbp 11.4

Data on the CHORI-242 clone average insert size has been determined by Pulsed Field Gel Electrophoresis. Clone size distribution 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.

  Ordering & Pricing information

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 ( ), fax: (510) 450-7924).




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