| RPCI
- 86 Male Feline BAC
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The RPCI-86 Male Feline BAC Library has been
constructed in our laboratory by Yu Wang. The project
was funded by Ralston Purina from a grant proposal by Leslie
A. Lyons, Ph.D., at The Laboratory of Genomic Diversity in
the National Cancer Institute. The library was made
from a random bred male domestic short-haired cat, a brown
classic tabby. Blood was collected and genomic DNA was
isolated from the white blood cells. The DNA (partially
EcoRI digested) was size selected and cloned between the EcoRI
sites of the pTARBAC2 vector. The ligation products
were transformed into DH10B electrocompetent cells (BRL LIFE
TECHNOLOGIES). The library has been arrayed into 384
well microtiter dishes and also gridded onto 22x22
cm high-density nylon hybridization filters for screening
by probe hybridization. Each hybridization membrane
represents over 18,000 distinct feline BAC clones, stamped
in duplicate
The Total RPCI-86 Feline Library:
| Segment |
Cloning
Vector |
DNA |
Plate Numbers |
Total Plates |
Total Clones |
Empty Wells (Total) |
| 1 |
pTARBAC2 |
Cat White Blood Cell |
1-288 |
288 |
107,041 |
781 |
| 2 |
pTARBAC2 |
Cat White Blood Cell |
289-624 |
336 |
127,308 |
1,320 |
| Total Library |
|
|
1-624 |
624 |
234,349 |
2,101 |
| Segment |
Empty Wells (%) |
Non-Recombinant Clones
(Total) |
Non-Recombinant Clones
(%) |
Insert Size (average) |
Genomic Coverage |
| 1 |
0.7 |
approx. 2,770 |
2.5 |
138 Kbp |
4.8x |
| 2 |
1.0 |
approx. 780 |
0.6 |
137 Kbp |
5.8x |
| Total Library |
0.9 |
approx. 3,550 |
1.5 |
137 Kbp |
10.6x |
click here
for a legend of the previous tables.
The
average insert size data has been determined by Pulsed
Field Gel Electrophoretic analysis of clones. Further
(in depth) characterization of the library is ongoing in our
lab and data will be updated on our WebPage periodically.
The library is available as library plates, individual clones, and high-density
hybridization sets (see Filter Availability)
Individual clones identified after screening can be obtained as stab cultures
at a cost of $80 per clone in addition to a $35 set-up charge per shipment. Shipping will be done by Federal Express and charged to the recipients account
number.
With respect to the scheduling of shipments of arrayed library copies, we would
like to make you aware of our policy for
array distribution.
Please visit the ordering information
page if you want to place an order.
Academic and commercial users interested in a copy of the BAC library should
contact Pieter J. de Jong (pdejong@chori.org, fax: (510) 450-7924). For hybridization
membranes, please contact BACPAC Resources (BACPACorders@chori.org).
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