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CHORI-26: Mouse CAST/Ei (M) (Mus musculus castaneus) BAC Library |
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The CHORI-26 Mus musculus castaneus (CAST/Ei) male mouse BAC Library has been constructed by Dr. 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. The mouse sample was obtained through the Jackson Laboratory. DNA was isolated from kidney cells of a Mus musculus castaneus (CAST/Ei) male mouse. Agarose-embedded high molecular weight DNA was partially digested with a combination of EcoRI restriction enzyme and EcoRI methylase. Size fractionated DNA was ligated into the pTARBAC2.1 vectors 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. For screening by probe hybridization, ten 22x22cm nylon high-density 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 Qing Cao and Kazutoyo Osoegawa. This work was funded by NIH grants HG01165-07SI and HG025323-01 as part of the NIH-funded BAC Resource Network .
Provisional data for CHORI-26 Mouse CAST/Ei 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 | kidney | kidney | kidney | Plate Numbers | 1-240 | 241-480 | 1-480 | Plate Count | 240 | 240 | 480 | Empty Wells | 1183 (1.28%) | 895 (0.97%) | 2078 (1.13%) | Non-Recombinant Clones | 0 (0%) | 0 (0%) | 0 (0%) | Non-Insert Clones | Approx. 1965 (5/232, 2.16%) | Approx. 2400 (6/228, 2.63%) | Approx. 4356 (11/460, 2.39%) | Recombinant Clones | 89012 | 88865 | 177886 | Average Insert Size | 170 Kbp | 170 Kbp | 170 Kbp | Genomic Coverage | 5X | 5X | 10X |
Click here for legend of the previous table.
No clones were found to be non-recombinant after analysis of CHORI-26 using overgo probes specific for the puc19 fragment.
Data on the CHORI-26 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 click here.
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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