| RPCI
- 22 (129S6/SvEvTac) Mouse BAC Library |
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The RPCI-22 BAC Library has been constructed in our laboratory
by Kazutoyo Osoegawa.
Female 129S6/SvEvTac (Taconic) mouse spleen genomic
DNA was isolated and partially digested with a combination
of EcoRI and EcoRI Methylase. Size selected DNA was
cloned into 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 - 22 129S6/SvEvTac Mouse BAC Library:
Segment |
Cloning
Vector |
DNA |
Plate Numbers |
Total Plates |
Total Clones |
Empty Wells (Total) |
| 1 |
pBACe3.6 |
Female Spleen |
1-288 |
288 |
102,914 |
1,678 |
| 2 |
pBACe3.6 |
Female Spleen |
289-576 |
288 |
109,039 |
1,443 |
| Total Library |
|
|
|
576 |
211,953 |
3,121 |
| Segment |
Empty Wells (%) |
Non-Recombinant Clones
(Total) |
Non-Recombinant Clones
(%) |
Average Insert Size |
Genomic Coverage |
| 1 |
1.5 |
approx. 6,000 |
5.5 |
155 Kbp |
5.3X |
| 2 |
1.3 |
approx. 110 |
0.1 |
154 Kbp |
5.6X |
| Total Library |
1.4 |
approx. 6,110 |
2.9 |
154 Kbp |
10.9X |
click here
for a legend of the previous tables.
Data on the RPCI-22 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 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 ([email protected], fax: (510) 450-7924). For hybridization
membranes, please contact BACPAC Resources ([email protected]).
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