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CHORI-233: Deer mouse (Peromyscus maniculatus rufinus) BAC Library |
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The CHORI-233 Deer Mouse (Peromyscus maniculatus rufinus) BAC library has been constructed by Boudewijn ten Hallers 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.
Kidney was obtained from a female Deer Mouse by Brian Hjelle (Department of Pathology, Infectious and Inflammation Program and Emerging Infectious Disease Centre) at the University of New Mexico School of Medicine. High-molecular-weight DNA was isolated from frozen kidney tissue by following the protocol developed in our laboratory 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 576 384-well microtiter dishes and will subsequently be gridded onto twelve 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).
Segment | 1 | 2 | All | Vector | pTARBAC2.1 | pTARBAC2.1 | pTARBAC2.1 | Restriction Enzyme | EcoRI/EcoRI Methylase | EcoRI/EcoRI Methylase | EcoRI/EcoRI Methylase | DNA Source | kidney | kidney | kidney | Plate Numbers | 1-288 | 289-576 | 1-576 | Plate Count | 288 | 288 | 576 | Empty Wells | 2828 (2.56%) | 2351 (2.13%) | 5179 (2.34%) | Non-Recombinant Clones | N/A | N/A | N/A | Non-Insert Clones | Approx. 1929 (5/280, 1.79%) | Approx. 1169 (3/279, 1.08%) | Approx. 3089 (8/559, 1.43%) | Recombinant Clones | 105835 | 107072 | 212916 | Average Insert Size | 180 Kbp | 185 Kbp | 182.5 Kbp | Genomic Coverage | 4.8X | 5X | 9.8X |
Data on the CHORI-233 clone insert size distribution has been determined by pulsed-field gel Electrophoresis. Clone Size Distribution has been plotted graphically.
No clones were found to be non-recombinant after analysis of CHORI-233 using overgo probes specific for the puc19 fragment.
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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