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CHORI-251: Chimpanzee (M) (Pan troglodytes) BAC Library |
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The CHORI-251 Chimpanzee BAC library has been constructed at the Children's Hospital Oakland Research Institute, BACPAC Resources, by Baoli Zhu for segments 1 and 2, Michael Nefedov for segments 3 and 4 using the cloning techniques developed in our laboratory (Osoegawa et al., 1998). DNA was isolated from white blood cells obtained from a male chimpanzee (Pan troglodytes, "Clint", Yerkes #C0471; birthdate: 6-6-80). Agarose embedded high molecular weight DNA was partially digested with a combination of EcoRI restriction enzyme and EcoRI Methylase for segments 1 and 2, and with MboI for segments 3 and 4. Size fractionated DNA was cloned into the pTARBAC2.1 vector between the EcoRI sites or pTARBAC1.3 vector between the BamHI sites. The ligation products were transformed into DH10B (T1 resistant) electrocompetent cells (BRL Life Technologies). The library has been arrayed into 384-well microtiter dishes and also gridded onto 24 22x22cm nylon high-density filter for screening by probe hybridization. Each hybridization membrane represents over 18,000 distinct Chimpanzee BAC clones, stamped in duplicate. BAC library construction was funded by a grant from NIH (#HG01165-06).
Segment | 1 | 2 | 3 | 4 | All | Vector | pTARBAC2.1 | pTARBAC2.1 | pTARBAC1.3 | pTARBAC1.3 |
| Restriction Enzyme | EcoRI/EcoRI Methylase | EcoRI/EcoRI Methylase | MboI | MboI | N/A | DNA Source | blood | blood | blood | blood | blood | Plate Numbers | 1-288 | 289-576 | 577-864 | 865-1152 | 1-1152 | Plate Count | 288 | 288 | 288 | 288 | 1152 | Empty Wells | 1630 (1.47%) | 1399 (1.27%) | 2295 (2.08%) | 2251 (2.04%) | 7595 (1.72%) | Non-Recombinant Clones | 552 (0.51%) | 653 (0.6%) | N/A | N/A | 1205 (0.28%) | Non-Insert Clones | Approx. 3923 (3.6%) | Approx. 6552 (6%) | Approx. 2762 (7/275, 2.55%) | Approx. 3954 (10/274, 3.65%) | Approx. 20000 (4.6%) | Recombinant Clones | 104487 | 101988 | 105535 | 104387 | 413568 | Average Insert Size | 164 Kbp | 176 Kbp | 190 Kbp | 180 Kbp | 178 Kbp | Genomic Coverage | 5.2X | 5.4X | 6.3X | 6X | 22.9X |
click here for a legend of the previous tables.
No clones were found to be non-recombinant after analysis of CHORI-251 using overgo probes specific for the puc19 fragment.
Data on the CHORI-251 clone insert size distribution has been determined by pulsed-field gel Electrophoresis. Clone Size Distribution has been plotted graphically.
The library is available as library plates, individual clones, and high-density hybridization sets (see Filter Availability)
Individual clones identified after screening can be obtained as stab cultures at a cost of $80 per clone in addition to a $35 set-up charge per shipment. Shipping will be done by Federal Express and charged to the recipients account number.
With respect to the scheduling of shipments of arrayed library copies, we would like to make you aware of our policy for array 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 hybridization membranes, please contact BACPAC Resources ([email protected]).
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