Elizabeth, Assuming the PCR conditions were controlled and you know reagents, method of preparation of the sample etc are ok, I'd be surprised if this technique wasn't the most sensitive of the two you've described-it seems strange then that the least sensitive method should give you a positive signal. My suggestion would be to check conditions of the PCR with a positive control, that cell cultures have been grown in antibiotic free medium for at least three passages (that includes selection agents such as neomycin/geneticin which have anti-microbial properties) and then recheck alongside another technique. We have used the Genprobe detection kit for a number of years and it works very successfully-however the PCR should remain the most sensitive. I hope this is useful. Regards Simon Simon Q.J. Rice Gene Expression Sciences Smithkline Beecham Pharmaceuticals, NFSP-N, Harlow, UK. et.kraus@baylordallas.edu on 22-Feb-1999 22:55 To: cyto-inbox cc: (bcc: Simon Q Rice) Subject: Mycoplasma Detection This message is being posted for a co-worker. Not necessarily flow-related but could be useful to the group: Does anyone have experience in mycoplasma detection in cell lines? Our dilemma: PCR results (ATCC Mycoplasma Detection Kit) are negative while confocal microscopy detects positive DAPI labeling in the cytoplasm of the same cell line. Any help would be appreciated. Elizabeth T. Kraus et.kraus@baylordallas.edu
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