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RPCI-93: Trypanosoma brucei BAC Library |
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The RPCI-93 Trypanosoma brucei TREU 927 BAC Library has been
constructed in our laboratory by Baohui Zhao in collaboration
with The Institute for Genomic Research (TIGR) and Sara Melville's
group at the University of Cambridge. T. brucei 927/10.1
agarose embedded DNA was partially digested with a combination
of EcoRI and EcoRI Methylase or DpnII (see table below).
Size selected DNA was cloned into the pBACe3.6 vector between the
EcoRI or Bam HI 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
T. brucei BAC clones, stamped in duplicate.
The RPCI-93 Trypanosoma brucei TREU 927 BAC Library:
| Segment |
Cloning
Vector | Cloning Enzyme |
DNA |
Plate Numbers |
Total Plates |
Total Clones |
Empty Wells (Total) |
| 1 |
pBACe3.6 |
EcoRI |
T. brucei 927/10.1 |
1-24 |
24 |
9,063 |
43 |
| 2 |
pBACe3.6 |
DpnII |
T. brucei 927/10.1 |
25-48 |
24 |
8,963 |
53 |
| Total Library |
|
|
|
1-48 |
48 |
17,996 |
96 |
| Segment |
Empty Wells (%) |
Non-Recombinant
Clones (Total) | Non-Recombinant Clones
(%) | Average Insert Size |
Genomic Coverage |
| 1 |
0.5 |
approx. 110 |
1.2 |
152 Kbp |
46X |
| 2 |
0.6 |
approx. 230 |
2.5 |
128 Kbp |
38X |
| Total Library |
0.5 |
approx. 340 |
1.5 |
141 Kbp |
84X |
click here
for a legend of the previous tables.
The total library should represent approximately 84-fold
total genomic representation.
The RPCI-93 clone average insert size has been determined by Pulsed Field
Gel Electrophoresis. Clone size distribution has been
plotted graphically.
The RPCI-93 T. brucei TREU BAC Library is available both
as library plates and high-density hybridization sets (see
Filter Availability).
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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