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CHORI-213: Threespine Stickleback (Gasterosteus aculeatus) BAC Library |
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The CHORI-213 Three-spine Stickleback (Gasterosteus aculeatus) BAC library has been constructed by Baoli Zhu in Pieter de Jong's Laboratory at the Children's Hospital Oakland Research Institute using the cloning technique developed in our laboratory (Osoegawa et al., 1998). DNA was isolated from blood obtained from Three-spine Sticklebacks by Dr. Chris Amemiya at Benaroya Research Institute of Virginia Mason. Agarose embedded high-molecular-weight DNA was partially digested with a combination of EcoRI restriction enzyme and EcoRI Methylase. Size fractionated DNA was cloned 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 384-well microtiter dishes and also gridded onto four 22x22cm nylon high-density filters for screening by probe hybridization. Each hybridization membrane represents over 18,000 distinct stickleback BAC clones, stamped in duplicate.
Library construction was supported through a sub-contract from a grant awarded to Dr. David Kingsley at Stanford University.
Provisional data for CHORI-213 Three-spine Stickleback BAC Library:
Segment | 1 | 2 | All | Vector | pTARBAC2.1 | pTARBAC2 | pTARBAC2.1 | Restriction Enzyme | EcoRI/EcoRI Methylase | EcoRI/EcoRI Methylase | EcoRI/EcoRI Methylase | DNA Source | blood | blood | blood | Plate Numbers | 1-192 | 193-288 | 1-288 | Plate Count | 192 | 96 | 288 | Empty Wells | 1386 (1.88%) | 575 (1.56%) | 1961 (1.77%) | Non-Recombinant Clones | 264 (0.36%) | N/A | N/A | Non-Insert Clones | Approx. 988 (1.34%) | N/A | N/A | Recombinant Clones | 71090 | 36289 | 108631 | Average Insert Size | 190 Kbp | N/A | 190 Kbp | Genomic Coverage | 19.3X | N/A | 19.3X |
Click here for legend of the previous table.
No clones were found to be non-recombinant after analysis of CHORI-213 using overgo probes specific for the puc19 fragment.
Data on the CHORI-213 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.
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 ([email protected], fax: (510) 450-7924).
Reference
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