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Human BAC (RPCI-11)

Human BAC (RPCI-11)

The RP11 Human Male BAC library was generated by the BACPAC Resource Center (BPRC) at the Children’s Hospital Oakland Research Institute by Kazutoyo Osoegawa. The work was funded by the National Human Genome Research Institute (NHGRI, NIH).

Male blood DNA was partially digested using a combination of EcoRI and EcoRI Methylase. The size selected DNA was then cloned into the pBACe3.6 vector). For Segment 5 the same DNA was partially digested with MboI. This was then size selected and ligated into the cloning vector pTARBAC1. The products were then transformed into E.coli DH10B. The clones have been arrayed into 384 well plates.

 

Source BioScience offers the complete clone set of 1440 384-well microtitre plates or individual clones, for research purposes only. We can also make subsets of your chosen clones from the collection. Please contact us to discuss your requirements.

Protocols / Clone Handling

Clones are streaked onto LB agar containing chloramphenicol (20µg/ml) and should be stored at 4°C on arrival.
Plates contain LB broth with 8% glycerol and the appropriate antibiotic. The plates are sent on Dry Ice, and should be stored at -70 °C.

To use: 

Restreak the clone onto the same agar type, in such a way as to obtain single colonies. Incubate at 37°C overnight.
Single colonies are then ready for DNA isolation, if required, using this protocol which although described for PAC clones also works for BAC clones.
Clones can be grown in LB broth + chloramphenicol, and glycerol added after growth to a concentration of 25% for long term storage at -70°C.

References

Subsequent publication:

For any publications arising from the use of these clones please acknowledge the originators of the library, Kazutoyo Osoegawa, Minako Tateno and Pieter deJong, and Source BioScience for providing the clones in any publication arising out of using this resource.

References:

  1. Osoegawa, K., Mammoser, A. G., Wu, C., Frengen, E., Zeng, C., Catanese, J. J., de Jong, P. J. (2001) A Bacterial Artificial Chromosome Library for Sequencing the Complete Human Genome, Genome Research, Vol. 11, Issue 3, 483-496, March 2001
  2. Osoegawa, K., Woon, P.Y., Zhao, B., Frengen, E., Tateno, M., Catanese, J.J, and de Jong, P.J. (1998) An Improved Approach for Construction of Bacterial Artificial Chromosome Libraries, Genomics 52, 1-8; Article # GE985423.

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