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CHORI-82: Canine Boxer (F) (Canis familiaris) BAC Library |
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The CHORI-82 Canine boxer (Canis familiaris) BAC library has been constructed by Dr. 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). A blood sample for this study was donated to the laboratory of Elaine Ostrander at the Fred Hutchinson Cancer Research Center, 1100 Fairview Ave N., D4-100, Seattle WA 98112 and transfered to Oakland. High-molecular-weight DNA was isolated following embedding of white blood cells in agarose. The agarose-embedded DNA was partially digested with a combination of EcoRI restriction enzyme and EcoRI methylase and size fractionated by pulsed-field electrophoresis. Size fractionated DNA fragments were ligated 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 eleven 22x22cm nylon high-density filters for screening by probe hybridization. Each hybridization membrane represents over 18,000 distinct BAC clones, stamped in duplicate.
Library characterization was performed by Qing Cao, Boudewijn ten Hallers and Dr. Kazutoyo Osoegawa.
BAC library construction was funded by NIH grant #HG025323-02, as part of the NIH-funded BAC Resource Network .
Provisional data for CHORI-82 Canine boxer (Canis familiaris) BAC Library:
Segment | 1 | 2 | All | Vector | pTARBAC2.1 | pTARBAC2.1 | pTARBAC2.1 | Restriction Enzyme | EcoRI/EcoRI Methylase | EcoRI/EcoRI Methylase | EcoRI/EcoRI Methylase | DNA Source | blood | blood | blood | Plate Numbers | 1-288 | 289-528 | 1-528 | Plate Count | 288 | 240 | 528 | Empty Wells | 3164 (2.86%) | 1613 (1.75%) | 4777 (2.36%) | Non-Recombinant Clones | N/A | N/A | N/A | Non-Insert Clones | N/A | N/A | N/A | Recombinant Clones | 107428 | 90547 | 197975 | Average Insert Size | 173 Kbp | 170 Kbp | 171.5 Kbp | Genomic Coverage | 5.5X | 4.5X | 10X |
Click here for legend of the previous table.
No clones were found to be non-recombinant after analysis of CHORI-82 using overgo probes specific for the puc19 fragment.
Data on the CHORI-82 clone insert size distribution has been determined by pulsed-field gel Electrophoresis. Clone Size Distribution has been plotted graphically.
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.
Ordering & Pricing information
The library is available in several formats. Individual clones, and high-density hybridization filter are obtainable. For ordering and shipping details, please view the ordering and pricing information ordering and pricing information page.
Academic and commercial users interested in a copy of the BAC library should contact Pieter J. de Jong ([email protected], fax: (510) 450-7924).
Reference
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