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BAC Library Construction

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   VMRC-6: Opossum (Monodelphis domestica) BAC Library   

The laboratory opossum BAC library was constructed by Andrew Stuart and Wendy Lippmann in Chris Amemiya’s laboratory at the Virginia Mason Research Center (VMRC). This library is designated VMRC-6. Blood was obtained from a male laboratory opossum (Monodelphis domestica) kindly provided by Dr. Paul Samollow, Southwest Foundation for Biomedical Research (San Antonio, Texas). Leucocytes 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 pCC1BAC vector (Epicentre Technologies). 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 grants 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 pCC1BAC Leucocyte EcoRI 1-336 336 N/A


2 pCC1BAC Leucocyte EcoRI 337-720 384 N/A N/A


pCC1BAC Leucocyte EcoRI 1-720 720 N/A N/A


Segment Ghost clones (including vector artifacts) Recombinant clones Insert Size (Average) Redundancy (Genome Size: 4200 Mb)


5% 95% 155 kb ~ 10X




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