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CHORI-270: Japanese macaque BAC Library |
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The CHORI-270 Japanese macaque (Macaca fuscata, male) BAC library has been constructed by Yuko Yoshonaga 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). A blood sample was obtained from a male Japanese macaque, "Wakasa". High-molecular-weight DNA was isolated from white blood cells by embedding in agarose. The embedded DNA was partially digested with a combination of EcoRI restriction enzyme and EcoRI Methylase and size fractionated by pulsed-field electrophoresis. DNA fragments from the appropriate size fraction were cloned into the pTARBAC2.1 vector between the two EcoRI sites. The ligation products were transformed into DH10B (T1 resistant) electro-competent cells (Invitrogen). The library has been arrayed into 480 “384-well” microtiter dishes and has subsequently been gridded onto ten 22x22cm nylon high-density filters for probe hybridization screening. Each hybridization membrane represents over 18,000 distinct BAC clones (48 dishes), stamped in duplicate. Library characterization was performed by Qing Cao, Teresa Ren and Kazutoyo Osoegawa.
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 | 1-240 | 241-480 | 1-480 | Plate Count | 240 | 240 | 480 | Empty Wells | 1935 (2.1%) | 1143 (1.24%) | N/A | Non-Recombinant Clones | N/A | N/A | N/A | Non-Insert Clones | Approx. 1244 (3/222, 1.35%) | Approx. 1889 (4/195, 2.05%) | N/A | Recombinant Clones | 88981 | 89128 | 184320 | Average Insert Size | 157 Kbp | 150 Kbp | N/A | Genomic Coverage | N/A | N/A | N/A |
Hybridization filters and individual clones can be ordered online. Filters can be obtained only per complete set of 10 membranes (price per set is 10 times $200.00). Shipping will be done by Federal Express and is preferably charged to the Fedex account number of the recipient. Information about cost and conditions are listed on our ordering instruction page.
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