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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 5-week old
female C57BL/6J mice (obtained from the Jackson Laboratory).
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 in the pBACe3.6
vector between 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 individual clone orders, distributed as agar-stab
cultures at a not-for profit cost. See our webpage "Ordering
& Pricing Information" for
the costs.
The BAC library has been gridded onto 22x22cm positively charged
nylon filters for hybridization screening purposes. Each filter
contains 36,864 colonies which represents 18,432 independent
clones spotted in duplicate in a 4x4 clone array. Clones can
be identified through screening by purchasing "high-density
colony" hybridization filters or by utilizing our "fee
for service" "Library Screening
Services" . For questions about clones and hybridization
membranes, please contact BACPAC Resources ([email protected]).
*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 ([email protected], fax: (510) 450-7924). For hybridization
membranes, please contact BACPAC Resources ([email protected]).
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