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RPCI-98: Drosophila melanogaster BAC Library |
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The RPCI-98 Drosophila melanogaster BAC Library has been
constructed in our laboratory by Aaron Mammoser and Kazutoyo
Osoegawa in collaboration with the Berkeley
Drosophila Genome Project headed by Gerald M. Rubin and
co-directed by Susan E. Celniker at Lawrence Berkeley National
Laboratory. Please note that this library has two different
names as a result of different nomenclature guidelines used
by the BGDP and CHORI (BACPAC Resources). The BGDP followed
the established nomenclature for various Drosophila
BAC libraries and BACPAC Resources followed the NCBI
recommendations. Hence, a clone called "BACR48M07"
by the BDGP should be referred to as "RP98-48M7".
High-molecular-weight DNA from an isogenic y1; cn1 bw1 sp1
strain (B.J. Brizuela et al., 1994, Genetics 137: 803; Bloomington
stock number 2057) was prepared by Roger A. Hoskins. D. melanogaster
agarose-embedded DNA was 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 a 22x22cm nylon
high-density filter for screening by probe hybridization.
The hybridization membrane represents over 18,000 distinct
D. melanogaster BAC clones, stamped in duplicate.
The RPCI-98 Drosophila melanogaster BAC Library:
| Segment |
Cloning Vector |
DNA |
Plate Numbers |
Total Plates |
Total Clones |
Empty Wells (Total) |
| Total Library |
pBACe3.6 |
Drosophila melanogaster |
1-48 |
48 |
17,540 |
243 |
| Segment |
Empty Wells (%) |
Non-insert clones |
pUC positive clones |
Non-Recombinant Clones (%) |
Average Insert Size |
Genomic Coverage |
| Total Library |
1.3 |
approx. 650 |
1 |
3.5 |
165 Kbp |
22X |
click here for a legend of the previous tables.
The total library should represent approximately 22-fold total genomic representation.
The RPCI-98 clone average
insert size has been determined by Pulsed Field Gel Electrophoresis.
Clone size distribution has been plotted graphically.
The RPCI-98 D. melanogaster BAC Library is available both
as library plates, individual clones, and high-density hybridization
sets (see Filter Availability)
Reference:
Hoskins, R. et al. (2000) A BAC-based physical map of the
major autosomes of Drosophila melanogaster. Science. 2000
Mar 24;287(5461):2271-4.
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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