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CHORI-234: Guinea pig (F) (Cavia porcellus) BAC Library |
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The CHORI-234 “guinea pig” (Cavia porcellus) BAC library has been constructed by Chung Li Shu and Kazutoyo Osoegawa 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). Snap frozen kidney was obtained at Covance Inc. derived from a highly inbred female guinea pig from the “2/N” breed (Covanche animal ID: B/P 213). The same animal is also being used as a DNA source for the Whole-Genome-Shotgun Sequencing at the Braod Institute. The frozen kidney has been ground under liquid nitrogen into fine powder using pestle and mortar. The powder was suspended in chromatin isolation buffer and further homogenized using a Dounce homogenizer with a tight pestle. The chromatin fragments were embedded in 0.5% agarose and treated with cell lysis buffer. The agarose embedded DNA was partially digested with a combination of EcoRI and EcoRI Methylase and size fractionated by pulsed-field electrophoresis. DNA fragments of the appropriate size fraction were cloned into the pBACGK1.1 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.
The BAC library construction was funded by grant (#HG025323-03) under the auspices of the NIH Resource Network.
Segment | 1 | 2 | All | Vector | pBACGK1.1 | pBACGK1.1 |
| Restriction Enzyme | EcoRI/EcoRI Methylase | EcoRI/EcoRI Methylase | N/A | DNA Source | kidney | kidney | N/A | Plate Numbers | 1-288 | 289-528 | 1-528 | Plate Count | 288 | 240 | 528 | Empty Wells | 3952 (3.57%) | 2699 (2.93%) | N/A | Non-Recombinant Clones | N/A | N/A | 19 (0.01%) | Non-Insert Clones | N/A | N/A | N/A | Recombinant Clones | 106640 | 89461 | 202733 | Average Insert Size | N/A | N/A | N/A | Genomic Coverage | N/A | N/A | N/A |
Click here for a legend of the previous tables.
No clones were found to be non-recombinant after analysis of CHORI-234 using overgo probes specific for the puc19 fragment. 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 recipient's account number.
Organized general distribution of clones and screening reagents remains exclusively the activity of BACPAC Resources, unless this service has ceased to exist.
All library copies and individual clones should be maintained using the recommended nomenclature guidelines as posted on the NCBI web site.
When the scientific results obtained from use of the CHORI-234 library are published or otherwise disseminated, the scientific creators and the origin of the BAC library should be acknowledged. A recommended reference will be posted on this page as soon as it is available. This is important to permit us to measure the impact of our scientific contributions and will facilitate continued funding of similar resources. Otherwise, please contact Pieter de Jong ([email protected]).
With respect to the scheduling of shipments of arrayed library copies, we would like to make you aware of our policy for array distribution.
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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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