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BAC Library Construction

 Home > Resources > Libraries
  CHORI-82 Canine BAC Library   
  Background information


The “CHORI-82” Canine 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). DNA was isolated from white blood cells obtained from a female dog named Tasha (breed: Boxer; family name: Canis familiaris). The sample was kindly provided by Gregory M. Acland in the Center for Canine Genetics & Reproduction, James A. Baker Institute for Animal Health, NYS College of Veterinary Medicine Cornell University, Hungerford Hill Rd, Ithaca, New York 14853 6401. High-molecular-weight DNA was isolated from white blood cells and was embedded in agarose to stabilize the DNA. The embedded DNA was partially digested with a combination of EcoRI restriction enzyme and EcoRI methylase and size fractionated by pulsed-field electrophoresis. DNA fragments from the appropriate size fraction were cloned into the pTARBAC2.1 vector between the two EcoRI sites. The ligation products were transformed into DH10B (T1 resistant) electro-competent cells (Invitrogen). The library will be arrayed into 528 384-well microtiter dishes and will subsequently be gridded onto 11 22x22cm nylon high-density filters for screening by probe hybridization. Each hybridization membrane represents over 18,000 distinct BAC clones, stamped in duplicate. BAC library construction was funded as part of the NIH-funded BAC Resource Network by a grant from NIH (#HG025323-02).

Provisional Data for CHORI-82 Canine BAC library:

Segment 1 2 Total
Cloning Vector pTARBAC2.1 pTARBAC2.1  
DNA Source White blood cells White blood cells  
Restriction Enzyme EcoRI EcoRI  
Plate Range 1-288 289-528 1-528
Total 384-Well Dishes 288 240 528
Empty Wells(%) N/A N/A N/A
Non-insert Clones (%) N/A N/A N/A
Vector Clones (%) N/A N/A N/A
Recombinant Clones N/A N/A N/A
Insert size (Average) ~185 Kbp ~185 Kbp ~185Kbp
Standard Deviation N/A N/A N/A
Library Redundancy (3.3 Gbp genome) ~5.5 fold ~4.5 fold ~10 fold

The CHORI-82 clone average insert size has been determined from a sample of 172 colonies prior to the arraying of the BAC library. Pulsed Field Gel Electrophoresis of Not1 digested BACs with appropriate size standard was used for size determination. Clone size distribution will be posted on this page as soon as available. 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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