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VMRC-6: Opossum (Monodelphis domestica) BAC Library |
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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 |
N/A |
| 2 |
pCC1BAC |
Leucocyte |
EcoRI |
337-720 |
384 |
N/A |
N/A |
All
|
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) |
All |
5% |
95% |
155 kb |
~ 10X |
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