All of the RPCI and CHORI genomic
DNA libraries have been gridded onto 22x22cm positively charged
nylon filters for hybridization screening purposes. Each filter
( Schleicher & Schuell Membrane Nytran -M pore size 0.45
µm) 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 or can be downloaded
from our download page ).
RPCI and CHORI 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."
clones are available, contact BACPAC
Resources (BACPACorders@chori.org) with requests.
Filters are available
only as complete library sets or segments at $200 per membrane.
Please check "Filter Set
Price List" for library filter set prices.
For shipping and
general ordering details, please view ordering
& pricing information page.