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CHORI-261: Chicken BAC Library |
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The CHORI-261 Chicken BAC Library has been constructed by Michael Nefedov 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). A whole blood sample from a chicken (jungle fowl, female) was kindly provided by Jerry Dodgson (Michigan State University). The animal (Jungle Fowl UCD001, Inbred 256 (female) used for the blood sample has also been used for three additional BAC libraries prepared at Texas A&M University and being used for the chicken genome sequencing project at Washington University in St.Louis. DNA was isolated from the whole blood by embedding a mixture of erythrocytes and lymphocytes in agarose. The agarose-embedded high-molecular weight DNA was partially-digested with a combination of EcoRI restriction enzyme and EcoRI Methylase. The DNA fragments were size-fractionated by pulsed field electrophoresis and DNA fragments from the appropriate size fraction were cloned into the pTARBAC2.1 vector between the EcoRI sites. The ligation products were then transformed into DH10B (T1 resistant) electro-competent cells (Invitrogen). The library has been arrayed into 192 [384-well] microtiter dishes. For screening by probe hybridization, four 22x22cm nylon high-density colony filters have been prepared. Each hybridization membrane represents over 18,000 distinct BAC clones from 48 microtiter dishes, represented by duplicate colonies. Library characterization (see below) was performed by Teresa Ren, Qing Cao, & Kazutoyo Osoegawa. This work was funded by grant support from the USDA under the auspices of the USDA-CSREES National Animal Genome Research Program (USDA Grant: #99-35205-8537).
| Cloning Vector | pTARBAC2.1 | | DNA Source | Whole blood | | Restriction Enzyme | EcoRI | | Plate range | (1 - 192) | | Total 384-well dishes | 192 | | Empty wells | 2,158 (-2.90%) |
| Non-insert clones | 0/190(<0.5%) | | Vector clones | 1/73k (<<1%) | | Recombinant Clones | ~ 73,000 (>97% of wells) | | Insert Size Average | ~ 182 kb | | Standard Deviation | 36 kb | | Library Redundancy | ~11 fold |
Click here for a legend of the previous tables.
Data on the CHORI-261 clone average insert size has been determined by Pulsed Field Gel Electrophoresis. Clone size distribution has been plotted graphically.
Hybridization filters can be ordered only per complete set of 4 filter membranes (price per set is 4 times $200). 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 of arrayed library copies 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 Pieter J. de Jong for questions related to this library.
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