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CHORI-280: Rock Hyrax (F) (Procavia capensis) BAC Library |
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The CHORI-280 “Rock Hyrax ” BAC library has been constructed by Boudewijn ten Hallers and Maxim Koriabine, Ph.D., in Pieter de Jong’s laboratory at BACPAC Resources, Children’s Hospital Oakland Research Institute. The preparation of the library followed the general cloning approach developed in our laboratory (Osoegawa et al., 1998).. DNA was isolated from kidney tissue from a female hyrax kindly provided by Lynne Nazareth at the Human Genome Sequencing Center, Department of Molecular & Human Genetics, Baylor College of Medicine, Houston, Texas. 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 has been arrayed into 528 384-well microtiter dishes and has subsequently been 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 by NIH grant HG025323-03 under the auspices of the NIH-funded BAC Resource Network.
Segment | 1 | 2 | All | Vector | pTARBAC2.1 | pTARBAC2.1 |
| Restriction Enzyme | EcoRI/EcoRI Methylase | EcoRI/EcoRI Methylase | N/A | DNA Source | kidney/brain | kidney/brain | N/A | Plate Numbers | 1-288 | 289-528 | 1-528 | Plate Count | 288 | 240 | 528 | Empty Wells | N/A | N/A | N/A | Non-Recombinant Clones | N/A | N/A | N/A | Non-Insert Clones | N/A | N/A | N/A | Recombinant Clones | 110592 | 92160 | 202752 | Average Insert Size | 198 Kbp | 161 Kbp | 182 Kbp | Genomic Coverage | N/A | N/A | N/A |
Click here for legend of the previous table.
No clones were found to be non-recombinant after analysis of CHORI-280 using overgo probes specific for the puc19 fragment.
Data on the CHORI-280 clone insert size distribution has been determined by pulsed-field gel Electrophoresis. Clone Size Distribution has been plotted graphically.
The library is available for individual clone orders distributed as agar-stab cultures in a fee structure described on our website "Ordering & Pricing Information" .
The BAC library has been gridded onto 22x22cm positively charged nylon filters for hybridization screening purposes. Each filter contains 36,864 colonies which represents 18,432 independent clones spotted in duplicate in a 4x4 clone array. Clones can be identified through screening by purchasing "high-density colony" hybridization filters or by utilizing our "fee for service" "Library Screening Services" . For questions about clones and hybridization membranes, please contact BACPAC Resources ([email protected]).
The library may also be available to academic users by reimbursement of duplication costs (labor & materials). Dishes can only be obtained per complete segment of the library (about half of the library) or by purchasing the entire library. Shipping will be done by Federal Express and charged to the recipients account number. With respect to the scheduling of shipments for arrayed library copies, we would like to make you aware of our policy for array library distribution. Please visit the ordering information page if you want to place an order. Academic and commercial users interested in a copy of the BAC library should contact Pieter J. de Jong ([email protected], fax: (510) 450-7924). For questions about hybridization membranes, please contact BACPAC Resources ([email protected]).
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