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[Arabidopsis] Re: Contamination of Arabidopsis seedlings

Allan via arab-gen%40net.bio.net (by allan.debono from gmail.com)
Thu May 31 10:58:25 EST 2012


On Tuesday, May 29, 2012 10:22:44 AM UTC-7, Eloísa Hernández Lucero wrote:
> Dear Colleagues,
> 
> I'm looking for some help. We are having serious problems with fungal
> contamination when working with two mutant lines of Arabidopsis. The
> seeds germinate well in MS medium, but after week starts
> contamination, specifically in mutant seeds because in the same plate
> are grown on one side wild seeds and are not contaminated. We tested
> sterilized seeds with ketoconazole, also a sterilization protocol with
> 70% ethanol solution prior to sterilization with chlorine and 1% SDS
> but so far we have not had good results. Any idea that could help to
> prevent this contamination?
> 
> Thank you very much,
> 
> Eloísa
> 
> 
> -- 
> Eloísa Hernández Lucero
> Departamento de Bioquímica L-103
> Facultad de Quimica Conjunto "E"
> UNAM

Hi Eloisa,

I've had good luck with a biocide called "PPM."
I started using PPM after a recommendation from our tissue culture technician.  

My protocol for sterilization is as follows:
70% ethanol for no more than 5 minutes.
Rinse in sterile water containing 0.02% PPM

For especially contaminated seeds (powdery mildew outbreak during seed set)
PPM is added directly to the 1/2 MS, agar medium as well.



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