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CHORI-271: Gibbon (Nomascus leucogenys) BAC Library |
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The CHORI-271 gibbon BAC Library has been constructed by Boudewijn ten Hallers and Baoli Zhu in Pieter de Jong’s laboratory at BACPAC Resources, Children’s Hospital Oakland Research Institute. The preparation of the library followed the cloning approach developed in our laboratory (Osoegawa et al., 1998). DNA was isolated from blood from a wild-born Northern White-cheeked female “Asia” with international studbook #0098. The blood was kindly provided by Alan Mootnick, Director of the Gibbon Conservation Center (GCC), Santa Clarita, California. Blood samples were collected on May 18th 2004. High-molecular-weight DNA was isolated from white blood cells by embedding in 0.5% agarose gel. 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 480 “384-well” microtiter dishes and has subsequently been gridded onto ten 22x22cm nylon high-density filters for probe hybridization screening. Each hybridization membrane represents over 18,000 distinct gibbon BAC clones (48 dishes), stamped in duplicate. Library characterization was performed by Teresa Ren and Dr. Kazutoyo Osoegawa. 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 | pTARBAC2.1 | Restriction Enzyme | EcoRI/EcoRI Methylase | EcoRI/EcoRI Methylase | EcoRI/EcoRI Methylase | DNA Source | WBC | WBC | N/A | Plate Numbers | 1-240 | 241-480 | 1-480 | Plate Count | 240 | 240 | 480 | Empty Wells | 3016 (3.27%) | 2080 (2.26%) | 5096 (2.76%) | Non-Recombinant Clones | N/A | N/A | N/A | Non-Insert Clones | Approx. 1605 (4/222, 1.8%) | Approx. 441 (1/206, 0.49%) | Approx. 2097 (5/428, 1.17%) | Recombinant Clones | 87539 | 89639 | 177127 | Average Insert Size | 171 Kbp | 175 Kbp | 173 Kbp | Genomic Coverage | N/A | N/A | N/A |
Click here for a legend of the previous tables.
Data on the CHORI-271 clone insert size distribution has been determined by pulsed-field gel Electrophoresis. Clone Size Distribution has been plotted graphically.
Hybridization filters can be ordered per complete set of 10 membranes (price per set is 10 times $200.00) or per segment (5 times $200.00). 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.
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]).
Please contact Pieter J. de Jong for questions related to this library.
Reference
Carbone L, Vessere GM, ten Hallers BF, Zhu B, Osoegawa K, Mootnick A, Kofler A, Wienberg J, Rogers J, Humphray S, Scott C, Harris RA, Milosavljevic A, de Jong PJ. A high-resolution map of synteny disruptions in gibbon and human genomes. PLoS Genet. 2006 2:e223.
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