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Mouse BAC (RPCI-23)

Mouse BAC (RPCI-23)

The RPCI-23 (RP23) BAC Library was constructed in Pieter de Jong's lab (Children's Hospital Oakland Research Institute) by visiting scientists Kazutoyo Osoegawa and Minako Tateno (RIKEN Tsukuba Life Science Center). From pooled tissues derived from three female C57BL/6J mice, kidney and brain genomic DNA samples were isolated and partially digested with a combination of EcoRI and EcoRI Methylase. Size-selected EcoRI fragments were cloned into the vector pBACe3.6 at the EcoRI sites. The ligation products were transformed into DH10B electro-competent cells (BRL Life Technologies).

450 384 well plates.

Protocols / Clone Handling

Clones are streaked onto LB agar containing chloramphenicol (20µg/ml).

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 the accompanying 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

Osoegawa K, Tateno M, Woon PY, Frengen E, Mammoser AG, Catanese JJ, Hayashizaki Y, de Jong PJ.Bacterial artificial chromosome libraries for mouse sequencing and functional analysis. Genome Res. 2000 Jan;10(1):116-28.


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