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CHORI-244: Indian muntjac (M) (Muntiacus muntjak) BAC Library

The CHORI-244 Indian muntjak BAC library has been constructed by Maxim Koriabine in Pieter de Jong’s laboratory at BACPAC Resources, Children’s Hospital Oakland Research Institute. The preparation of the library followed the general cloning approach developed in our laboratory (Osoegawa et al., 1998). DNA was isolated from a sample obtained from the San Diego Zoo for a male North Indian Muntjak (Muntiacus muntjak vaginalis; San Diego Zoo Accession # M0697034; Born 01/23/1997). 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 576 “384-well” microtiter dishes and has subsequently been gridded onto twelve 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. BAC library construction was funded by NIH grant HG025323-02 under the auspices of the NIH-funded BAC Resource Network.

Web page updated: August 27, 2005