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CHORI-87: Clouded leopard (Neofelis nebulosa) BAC Library |
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The CHORI-87 Clouded leopard (Neofelis nebulosa) BAC library has been constructed by Dr. Baoli Zhu in Pieter de Jong's laboratory at BACPAC Resources, Children’s Hospital Oakland Research Institute. Library preparation followed the general cloning approach developed in our laboratory Osoegawa et al., 1998.
DNA was isolated from white blood cells obtained from the National Zoo Conservation & Research Center in Front Royal, VA. Agarose-embedded high molecular weight DNA was partially digested using a combination of EcoRI restriction enzyme and EcoRI methylase. Size fractionated DNA fragments were ligated into the pTARBAC2.1 vector between the EcoRI sites. The ligation products were transformed into DH10B(T1 resistant)electrocompetent cells (Invitrogen).
The library has been arrayed into 480, 384-well microtiter dishes and subsequently gridded onto 10, 22x22cm nylon membranes. The 22x22cm nylon high-density filters have been made available for probe hybridization screening.
Library characterization was performed by Teresa Ren and Dr. Kazutoyo Osoegawa. BAC library construction was funded by grant (#HG025323-03) under the auspices of the NIH Resource Network.
Provisional data for CHORI-87 Clouded leopard BAC Library:
Segment | 1 | 2 | All | Vector | pTARBAC2.1 | pTARBAC2.1 | pTARBAC2.1 | Restriction Enzyme | EcoRI/EcoRI Methylase | EcoRI/EcoRI Methylase | EcoRI/EcoRI Methylase | DNA Source | N/A | N/A | N/A | Plate Numbers | 1-240 | 241-480 | 1-480 | Plate Count | 240 | 240 | 480 | Empty Wells | 2485 (2.7%) | 3247 (3.52%) | 5732 (3.11%) | Non-Recombinant Clones | 11 (0.01%) | 39 (0.04%) | 50 (0.03%) | Non-Insert Clones | Approx. 0 (0/220, 0%) | Approx. 1485 (4/239, 1.67%) | Approx. 1554 (4/459, 0.87%) | Recombinant Clones | 89664 | 87389 | 176984 | Average Insert Size | N/A | N/A | 195 Kbp | Genomic Coverage | N/A | N/A | N/A |
Click here for legend of the previous table.
No clones were found to be non-recombinant after analysis of CHORI-87 using overgo probes specific for the puc19 fragment.
Data on the CHORI-87 clone insert size distribution has been determined by pulsed-field gel Electrophoresis. Clone Size Distribution has been plotted graphically.
Further in depth characterization of the library is on going in our lab and data will be updated on our web page periodically.
Please direct questions concerning this library to either Pieter J. de Jong or Kazutoyo Osoegawa .
Ordering & Pricing information
The library is available in several formats. Individual clones, and high-density hybridization filter are obtainable. For ordering and shipping details, please view the ordering and pricing information ordering and pricing information page.
Academic and commercial users interested in a copy of the BAC library should contact Pieter J. de Jong (pdejong@chori.org, fax: (510) 450-7924).
Reference
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