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 High-Density Hybridization Filters    

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 (BACPACorders@chori.org) 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 (BACPACorders@chori.org) for further information regarding current availability or ordering.

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