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RPCI - 23 Female (C57BL/6J) Mouse BAC Library
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The RPCI-23 BAC Library has been constructed in our laboratory
by Kazutoyo Osoegawa and Minako Tateno* from pooled tissues
derived from three female C57BL/6J mice. Mouse kidney and
brain genomic DNA samples were isolated and partially digested
with a combination of EcoRI and EcoRI Methylase. Size-selected
EcoRI fragments were cloned between the pBACe3.6 vector at
the EcoRI sites. 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 22x22cm
nylon high-density filters for screening by probe hybridization.
Each hybridization membrane represents over 18,000 distinct
mouse BAC clones, stamped in duplicate.
The RPCI-23 C57BL/6J Mouse BAC Library:
| Segments |
Cloning
Vector |
DNA |
Plate Numbers |
Total Plates |
Total Clones |
Empty Wells (Total) |
| 1 |
pBACe3.6 |
Female (kidney/brain) |
1-240 |
240 |
90,443 |
1717 |
| 2 |
pBACe3.6 |
Female (kidney/brain) |
241-480 |
240 |
89,953 |
2207 |
| Total Library |
|
|
|
480 |
180,396 |
3924 |
| Segments |
% Empty Wells |
Non-Recombinant Clones (Total) |
Non-Recombinant Clones (%) |
Insert Size (average) |
Genomic Coverage |
| 1 |
1.9 |
approx. 10523 |
11.3 |
198 Kbp |
5.3X |
| 2 |
2.4 |
approx. 1211 |
1.3 |
197 Kbp |
5.9X |
| Total Library |
2.1 |
approx. 11734 |
6.8 |
197 Kbp |
11.2X |
click here for a legend of
the previous tables.
Data on the RPCI-23 clone average
insert size has been determined by Pulsed Field Gel Electrophoresis.
Clone size distribution has been plotted graphically.
Reference:
Osoegawa, K. et al. (2000) Bacterial Artificial Chromosome
Libraries for Mouse Sequencing and Functional Analysis.
Genome Research 10: 116-128
Reprints availiable upon request.
The library is available for academic users by reimbursement of duplication
costs (labor & materials), resulting in a price of $20 per 384-well dish.
Dishes can only be obtained per complete library
($9,600)
or per segment
($4,800)Individual clones identified after screening can be obtained as stab cultures
at a cost of $25 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..
*Visiting scientist supported by the RIKEN Institute, permanent
address:
Minako Tateno
RIKEN Tsukuba Life
Science Center
Genome Science
Laboratory
3-1-1 Koyadai Tsukuba,
Ibaraki 305 Japan
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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