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VMRC-15: African elephant (M) (Loxodonta africana) BAC Library |
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The African elephant (Loxodonta africana) BAC library was constructed by Michael Schoenborn in Chris Amemiya’s laboratory at the Benaroya Research Institute at Virginia Mason (BRI). This library was designated VMRC-15. A freshly taken blood sample was obtained from a male African elephant from Disney’s Animal Kingdom (from Barbara Burkhalter, Orlando, Florida). The particular animal (named “Macclean”) was wild-born around 1982, presumably in Namibia and has been registered in the North American Studbook for African elephants as #398. Leucocytes were separated via centrifugation, embedded in Incert agarose, and high molecular weight DNA was prepared in situ in the agarose. Generation of the library followed the general cloning approach developed in Pieter de Jong’s laboratory (Osoegawa et al., 1998). . EcoRI fragments from the appropriate size fraction were cloned into the EcoRI site of the pCCBAC vector (Epicentre Technologies). The ligation products were then 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. Analysis of 150 randomly selected BACs via pulsed field gel electrophoresis of NotI-digested DNA indicated that the average insert size was around 142 kb. The BAC library construction was funded by NIH grant HG02526 (Chris T. Amemiya, BRI) under the auspices of the NIH Resource Network.Segment | 1 | All | Vector | pCC1BAC | pCC1BAC | Restriction Enzyme | EcoRI/EcoRI Methylase | EcoRI/EcoRI Methylase | DNA Source | WBC | N/A | Plate Numbers | 1-600 | 1-600 | Plate Count | 600 | 600 | Empty Wells | 0 (0%) | 0 (0%) | Non-Recombinant Clones | 4 (0%) | 4 (0%) | Non-Insert Clones | N/A | N/A | Recombinant Clones | 230396 | 230396 | Average Insert Size | 142 Kbp | 142 Kbp | Genomic Coverage | N/A | 10X |
click here for a legend of the previous tables.
The library is available for individual clone orders distributed as agar-stab cultures at a cost described elsewhere. Clones can be identified through screening of "high-density colony" hybridization filters . The library may also be available to academic users by reimbursement of duplication costs (labor & materials), at a price of $20 per 384-well dish. Dishes can only be obtained per complete library. 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]).
Posted: November 30, 2004.
Revised:
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