|
RPCI-42: Male Bovine BAC Library |
 |
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 ([email protected], fax: (510) 450-7924). For hybridization membranes, please contact BACPAC Resources ([email protected]).
|
