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BAC Library Construction

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  VMRC-7 Bat BAC Library   

The bat BAC library was constructed by Alicia Hill-Force, Tsutomu Miyake and Joshua Danke in Chris Amemiya’s laboratory at the Virginia Mason Research Center (VMRC). This library was designated VMRC-7. Brain tissue was obtained from a male Greater Horsehoe Bat (Rhinolophus ferrumequinum) kindly provided from Dr. Walter Metzner, University of California, Los Angeles, CA. 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). DNA fragments from the appropriate size fraction were cloned into the pBAC-VM4 vector (GenBank no. pending). The ligation products were then transformed into DH10B (T1 resistant) electro-competent cells (Invitrogen). The library has been arrayed into 384-well microtiter dishes. This work was funded by NIH grant HG02526-01 (Chris T. Amemiya, VMRC).
Please note: The following data is provisional pending further characterization.

Segment Cloning Vector DNA Restriction enzyme Plate Number Total Plates Empty Wells Empty Wells (%)
1 pBAC-VM4 Brain EcoRI 1-402 402 0 0


Segment Ghost clones (including vector artifacts) Recombinant clones Insert Size (Average) Redundancy (Genome Size: 2300 Mb)
1 5% 95% 165 kb >= 10X




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