| RPCI
- 42 Male Bovine BAC Library
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The RPCI-42 Bovine Library has been constructed in our laboratory
By Chung-Li Shu with funding provided through the Monsanto
Corporation. Holstein Bull white blood cell 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
bovine BAC clones, stamped in duplicate.
The RPCI-42 Bovine BAC Library:
| Segment |
Cloning
Vector |
DNA |
Plate Numbers |
Total Plates |
Total Clones |
Empty Wells (Total) |
| 1 |
pBACe3.6 |
Holstein Bull White Blood
Cell |
1-288 |
288 |
108,776 |
1,156 |
| 2 |
pBACe3.6 |
Holstein Bull White Blood
Cell |
289-576 |
288 |
107,663 |
1,379 |
| Total Library |
|
|
|
576 |
216,439 |
2,535 |
| Segment |
Empty Wells (%) |
Non-Recombinant Clones
(Total) |
Non-Recombinant Clones
(%) |
Average Insert Size |
Genomic Coverage |
| 1 |
1.0 |
approx. 660 |
0.6 |
165 Kbp |
6.0X |
| 2 |
1.2 |
approx. 1,550 |
1.4 |
163 Kbp |
5.9X |
| Total Library |
1.1 |
approx. 2,210 |
1.0 |
164 Kbp |
11.9X |
click here
for a legend of the previous tables.
The total library should represent approximately 12-fold
total genomic representation. The RPCI-42 Bovine BAC
Library clone
average insert size has been determined by Pulsed Field
Gel Electrophoresis. Clone size distribution has been
plotted graphically. The RPCI-42 Bovine BAC Library
is available both as library plates and high-density hybridization
filter 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 (pdejong@chori.org, fax: (510) 450-7924). For hybridization
membranes, please contact BACPAC Resources (BACPACorders@chori.org).
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