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Help propogating Chlamydia stock

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#1 nube



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Posted 04 July 2012 - 03:13 PM

Hi All,

I am about to start trying to culture Chlamydia (C. pecorum and C. trachomatis, in BGM cells), and have never done any cell culture work before.

I have a few 100uL aliquots of C. trachomatis stock that I need to grow up in order to make 100's of aliquots, to use as my positive control. Each aliquot is apparently enough to inoculate one shell vial. Does anybody have a protocol for how to amplify the Chlamydia? Do I just grow up one of the aliquots in a shell vial, and after incubation use the supernatant to inoculate further shell vials, until I have a great enough volume? Or do I need to disrupt the cells somehow to release the EB's before harvesting the supernatant?

I'm sorry if this is a very basic question, but I am struggling to find a protocol for doing this. Any help will be greatly appreciated!

Thank you!

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