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BAC library screening
BAC Filters available for screening:
RPCI-11 BAC library, Segment 1 (Human male)
6 filter set, 5.8-fold genomic coverage
Average insert size, 164 Kbp
RPCI-22 BAC library, Segment 1 (Mouse 129S6/SvEvTac Female)
6 filter set, 5.3-fold genomic coverage
Average insert size, 155 Kbp
RPCI-23 BAC library, Segment 1 (Mouse C57BL/6J Female)
5 filter set, 5.3-fold genomic coverage
Average insert size, 198 Kbp
RPCI-24 BAC library, Segment 1&2 (Mouse C57BL/6J Male)
12 filter set, 10.8-fold genomic coverage
Average insert size, 155 Kbp
BAC Library Screening and Characterization - Overview
All of the RPCI genomic DNA libraries have been gridded onto 22x22cm positively charged nylon filters for hybridization screening purposes (see screening protocol). Each filter contains 36,864 colonies which represents 18,432 independent clones that have been spotted in duplicate in a 4x4 clone array. Example of RPCI-23 filter hybridized with 36 separate overgos.
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 below:
List of non recombinant clones:
RPCI-11 Human Male BAC Library
RPCI-13 Female Human BAC Library
RPCI-22 Female (129S6/SvEvTac) Mouse BAC Library
RPCI-23 Female (C57BL/6J) Mouse BAC Library
RPCI-24 Male (C57BL/6J) Mouse BAC Library
RPCI-32 Male (Brown Norway) Rat BAC Library
RPCI-41 Male (Olive) Baboon BAC Library
RPCI-42 Male Bovine BAC Library
RPCI-43 Male Chimpanzee BAC Library
RPCI-44 Male Porcine BAC Library
RPCI-81 Male (Doberman Pinscher) Canine BAC Library
RPCI-86 Male Feline BAC Library
RPCI-104 Schizosaccharomyces pombe Library
CHORI-14 Human male BAC Library
BAC LIBRARY SCREENING F.A.Q.s
Q. What can we do for you?
A. Using a custom-designed overgo generated from a sequence provided by you or a specific nucleic acid probe, we will screen the BAC library you specify and locate clones homologous to your probe. We usually locate 3-8 clones per unique probe.
Q. What other services do you offer?
A. We can isolate a single clone(s) from a library address and provide BAC glycerol stocks, agar stabs and/or DNA preps. Up to three clones are included for distribution as part of the BAC screening service. Additional clones are available for purchase as described on the fee schedule.
Q. What is an overgo probe?
A. An efficient methodology for isolation of BAC clones was developed by J. McPherson at
Q. Why are overgos preferred over other DNA probes?
A. The overgos designed are free of all known repetitive elements, E. Coli sequences, and vector sequences that would produce false-positive and non-specific hybridization-positive clones. The similar characteristics of each overgo designed also allows pooling of up to 50 overgos in a single hybridization experiment.
Q. Can we provide our own DNA probes?
A. Yes, we can use cDNAs, PCR products, cloned genomic DNA, synthetic oligonucleotides, and end probes from BACs. We need about 100 ng of purified probe for labeling (for most probes). The probe should be free of contaminating vector, PCR primers, and other nucleic acids that may create non-specific signals. For oligonucleotide probes, we need about 250 ng for labeling.
Q. How long does it take?
A. In general, 2-3 weeks from receipt of probe to shipping of clones. We begin a hybridization experiment the first Monday of each month.
Q. Is the service guaranteed?
A. Yes. If no clones are located using your probe, we do not charge for the screening, only the probe generation.
Q. How much does it cost?
A. The current cost of basic screening can be found on the Fee schedule and varies with the size of the BAC library and customer affiliation.
