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CHORI-262: California condor (F) (Gymnogyps californianus) BAC Library |
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The CHORI-262 “California Condor” BAC library has been constructed by Maxim Koriabine 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 1 ml of condor blood obtained at the San Diego Zoo as part of routine health monitoring. The sample was derived from a female condor ("Molloko" ISIS # 888071; studbook #45) and was kindly provided by Dr.Oliver Ryder. This female was the first captive bred condor at the San Diego Zoo, an F1 from parents (#13 & #20). Blood was mixed with agarose to embed the red and white blood cells. After solidifying the agarose-embedded cells in a plug mold, the plugs were then treated with detergents and proteinases as previously described. The agarose-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 384-well microtiter dishes and has subsequently been gridded onto 22x22cm nylon high-density colony filters for screening by probe hybridization. Each hybridization membrane represents over 18,000 distinct BAC clones (equivalent to 48 "384-well" dishes), stamped in duplicate.
Segment | 1 | All | Vector | pTARBAC2.1 |
| Restriction Enzyme | EcoRI/EcoRI Methylase | N/A | DNA Source | whole blood | N/A | Plate Numbers | 1-240 | 1-240 | Plate Count | 240 | 240 | Empty Wells | 2495 (2.71%) | 2495 (2.71%) | Non-Recombinant Clones | N/A | N/A | Non-Insert Clones | N/A | N/A | Recombinant Clones | 89665 | 89665 | Average Insert Size | N/A | N/A | Genomic Coverage | N/A | N/A |
References:
This BAC library was described in a publication comparing chicken and condor genes. See: Romanov M.N. et al, 2006; Construction of a California condor BAC library and first-generation chicken-condor comparative physical map as an endangered species conservation genomics resource
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