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CHORI-16: Human (Homo sapiens) Sheared BAC Library |
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The CHORI-16 library was generated by Kazutoyo Osoegawa. The preparation of the library followed the cloning approach developed in our laboratory (Osoegawa et al., submitted). This library was generated according to the 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 sheared by repeating freeze and thaw cycles. The fragmented ends were polished by T4 DNA polymerase. The polished ends were ligated to the blunt-end side of an adapter which has a 3'overhang. Size fractionated DNA was cloned into the pTARBAC6 vector between the BstXI sites. The ligation products were transformed into DH10B T1 phage resistant electrocompetent cells (Invitrogen). The library has been arrayed into 422 (384-well) microtiter dishes. The 22x22cm nylon high-density filters have been made available for probe hybridization screening.
The work was funded by grants from the National Institutes of Health (1RO1HL55700), the National Human Genome Research Institute (1RO1RG01165) and the US Department of Energy (DE-FG02-94ER61883).
Segment | 1 | All | Vector | pTARBAC6 | pTARBAC6 | Restriction Enzyme | sheared | sheared | DNA Source | N/A | N/A | Plate Numbers | 1-422 | 1-422 | Plate Count | 422 | 422 | Empty Wells | 3340 (2.06%) | 3340 (2.06%) | Non-Recombinant Clones | N/A | N/A | Non-Insert Clones | Approx. 7650 (20/415, 4.82%) | Approx. 7650 (20/415, 4.82%) | Recombinant Clones | 151058 | 151058 | Average Insert Size | 84 Kbp | 84 Kbp | Genomic Coverage | 4.1X | 4.1X |
Click here for legend of the previous table.
Data on the CHORI-16 clone insert size distribution has been determined by pulsed-field gel Electrophoresis. Clone Size Distribution has been plotted graphically.
Clone size distribution has been plotted graphically. While analyzing clones using pulse-field electrophoresis to determine the average insert size, non-insert clones containing a small deleted vector fragment consistent with sucrose resistance were observed. Further in depth characterization of the library is on going in our lab and data will be updated on our web page periodically.
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
Osoegawa K, Vessere GM, Li Shu C, Hoskins RA, Abad JP, de Pablos B, Villasante A, de Jong PJ. BAC clones generated from sheared DNA. Genomics 2007 89:291-9.
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