| RPCI
Human PAC Library
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RPCI Human PAC libraries have been
constructed in our laboratory by Panos A. Ioannou* (RPCI-1),
Chira Chen and Baohui Zhao (RPCI-3), and Baohui Zhao (RPCI-4,
5, and 6). Library construction was funded by grants
form the U.S. Department of Energy (#DE-FG02-94ER61883) and
the National Human Genome Research Institute (NHGRI, NIH)
(#1R01RG01165). Anonymous male (RPCI-1, 3, 4, AND 5)
or female (RPCI-6) blood DNA was isolated, partially digested
with MboI and cloned between the BamH1 sites of the pCYPAC2 (RPCI-1, 2, 3, 4, and 5) or pPAC4 (RPCI-6) vector. The ligation products were
transformed into DH10B electrocompetent cells (BRL Life technologies).
The libraries have been arrayed into 384-well microtiter dishes
and gridded onto 22x22cm nylon high
density hybridization filters for screening purposes.
The procedure for preparing the male PAC library including
the male PAC library, have been described by Ioannou
et al 1994. The data in this publication refers to the
first set of PAC clones (the RPCI-1 PAC library). The later
clone sets (RPCI-3, RPCI-4 and RPCI-5) were prepared by precisely
the same method using the same donor DNA and pCYPAC2 vector.
*Present Address:
Panos Ioannu, Ph.D.
The Murdoch Institute
for Research into Birth Defects
Royal Children's
Hospital
Flemington Road,
Parkville 3052
Melbourne, Australia
The RPCI Human PAC
Library is available directly
from Children's Hospital Oakland Research Institute. The Library
segments have been characterized as to:
- The percentage of empty wells
- Percentage of non-recombinant
clones
- Average insert size
- Approx. genomic coverage
The clone
size distribution for each segment of the library
has also been determined.
The library is
available for individual clone orders, distributed as agar-stab
cultures at a not-for profit cost. See our webpage "Ordering
& Pricing Information" for
the costs .
The PAC libraries have been gridded onto 22x22cm positively
charged nylon filters for hybridization screening purposes.
Each filter contains 36,864 colonies which represents 18,432
independent clones spotted in duplicate in a 4x4 clone array.
Clones can be identified through screening by purchasing "high-density
colony" hybridization filters or by utilizing our "fee
for service" "Library Screening
Services" . For questions about clones and hybridization
membranes, please contact BACPAC Resources (BACPACorders@chori.org).
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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