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VMRC-20: Ground squirrel BAC Library

The 13-lined ground squirrel (Spermophilus tridecemlineatus) BAC library was constructed by Andrew Stuart and Deb Tinnemore in Chris Amemiya’s laboratory at the Benaroya Research Institute at Virginia Mason (BRI). This library was designated VMRC-20. Brain tissue was obtained by Dr. Tom Miyake from a female ground squirrel from a colony housed at NIH (NINDS; lab of Dr. John Hallenbeck). Brain cells were embedded in Incert agarose and high molecular weight DNA was prepared in situ in the agarose. Generation of the library closely followed the cloning approach developed in Pieter de Jong’s laboratory Osoegawa et al., 1998. Partial-digest EcoRI fragments from the appropriate size fraction were cloned into the pCCBACE1 vector (Epicentre Technologies). The ligation products were then transformed into DH10B (T1 resistant) electro-competent cells (Invitrogen). The library has been arrayed into 720 384-well microtiter dishes and has subsequently been gridded onto 22x22cm nylon high-density colony filters for screening by probe hybridization. Each hybridization membrane represents over 18,000 distinct BAC clones (equivalent to 48 "384-well" dishes), stamped in duplicate. Analysis of 120 randomly selected BACs via pulsed field gel electrophoresis of NotI-digested DNA indicated that the average insert size was around 160 kb. The BAC library construction was funded by NIH grant HG02526 (Chris T. Amemiya, BRI) under the auspices of the NIH Resource Network.
Please note: The following data is provisional pending further characterization.