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CHORI-14: Human (M) (Homo sapiens) BAC Library |
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The CHORI-14 library was generated by Chung Li Shu and Dr. Kazutoyo Osoegawa. The preparation of the library followed the cloning approach developed in our laboratory Osoegawa et al., 1998. This library was generated according to the new NHGRI/DOE "Guidance on Human Subjects in Large-Scale DNA Sequencing". Male blood was obtained via a double-bind selection protocol. Male blood DNA was isolated from one randomly chosen donor (out of 10 male donors) and partially digested with a combination of EcoRI and EcoRI Methylase. Size selected DNA was cloned into the pTARBAC2 vector between the EcoRI sites. The ligation products were transformed into DH10B electrocompetent cells (BRL Life Technologies). The library has been arrayed into 528 (384-well) microtiter dishes. The 22x22cm nylon high-density filters have been made available for probe hybridization screening.
BAC library construction was funded by a grant from the National Human Genome Research Institute (NHGRI, NIH) (#HG01165-06).
Provisional data for CHORI-14 Human (male) 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 | blood | blood | blood | Plate Numbers | 1-282 | 289-528 | 1-528 | Plate Count | 282 | 240 | 528 | Empty Wells | 2694 (2.49%) | 2147 (2.33%) | 4841 (2.39%) | Non-Recombinant Clones | 24 (0.02%) | 42 (0.05%) | 66 (0.03%) | Non-Insert Clones | Approx. 4963 (4.7%) | Approx. 2610 (2.9%) | Approx. 7323 (3.7%) | Recombinant Clones | 100607 | 87361 | 190522 | Average Insert Size | 163 Kbp | 168 Kbp | 165.4 Kbp | Genomic Coverage | 5X | 4.5X | 9.5X |
Click here for legend of the previous table.
No clones were found to be non-recombinant after analysis of CHORI-14 using overgo probes specific for the puc19 fragment.
Data on the CHORI-14 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 complete copy of this BAC library is not available. The library is available in several formats. Individual clones, and high-density hybridization filter are obtainable. For ordering and shipping details, please click here.
Reference
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