| High-Density
Hybridization Filters |
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All of the RPCI genomic DNA 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
that have been spotted in duplicate in a 4x4 clone array.
Filter sets are available
directly from BACPAC Resources, Children's
Hospital Oakland Research Institute.
Filters should be
hybridized with (32)P-labeled (random-primed) probes (1x106 cpm/ml hybridization solution) using
standard conditions. After washing
to remove unbound probe, filters are wrapped in plastic film
(Saran) and exposed to x-ray film (Kodak Xar, Amersham Hyperfilm)
for 2 to 24 hrs. Complete documentation on interpretation
of hybridization results (location of positive clone plate
coordinates) is included with the filter sets).
Positive individual clones are available. Please refer to
the ordering information
page for pricing and shipping information. Please contact
BACPAC Resources ([email protected])
with requests.
The
RPCI BAC and PAC genomic libraries have varying amounts of
non-recombinant clones. A very small percentage of these
non-recombinant clones are the result of vector molecules
not cutting with the cloning enzyme. These clones do
not contain inserts as they still contain the pUC-link stuffer
fragment. They therefore hybridize very strongly to
probes containing pUC-like sequences. The locations
of these "pUC-Positive" clones can be found by clicking
on the hyperlink on the home page titled "Non-Recombinant Clones."
Email to BACPACorders ([email protected])
for further information regarding current availability or
ordering.
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