| RPCI-24
Mouse (C57BL/6J Male) BAC Library |
 |
The RPCI-24 BAC library has been
constructed in our laboratory from tissues derived from single
male C57BL/6J mouse (obtained from the Jackson Laboratory).
Spleen and brain genomic DNA samples were isolated and partially
digested with MboI. Size-selected MboI fragments were cloned
into the pTARBAC1 vector between the BamHI 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-24 Male
(C57BL/6J) Mouse BAC Library
| Segments |
Cloning
Vector |
DNA |
Restriction
enzyme |
Plate Numbers |
Total Plates |
Empty Wells |
Empty Wells
(%) |
| 1 |
pTARBAC1 |
Spleen |
MboI |
1-161, 168-177
|
288 |
3464 |
3.1 |
| Brain |
MboI |
162-167, 178-288
|
| 2 |
pTARBAC1 |
Brain |
MboI |
289-313 |
288 |
3951 |
3.6 |
| Spleen |
MboI |
314-576 |
| Total Library |
|
|
|
|
576 |
7415 |
3.35 |
| Segments |
Non-insert Clones (%) |
Non-insert clones |
Non-Recombinant
Clones (pUC) |
Recombinant Clones |
Average Insert Size |
Redundancy
(Genome size:
3,000 Mb) |
| 1 |
4.3 |
4607 |
1 |
102,521 |
160 Kbp |
5.5 |
| 2 |
0.5 |
533 |
0 |
106,108 |
150 Kbp |
5.3 |
| Total Library |
2.4 |
5140 |
1 |
208,629 |
155 Kbp |
10.8 |
click here
for a legend of the previous tables.
Data on the RPCI-24 clone average insert size (Segment
1, Segment
2) has been determined by Pulsed Field Gel Electrophoresis.
Clone size distribution has been plotted graphically.
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]).
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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