Contamination of Arabidopsis seedlings

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Contamination of Arabidopsis seedlings

Eloísa Hernández Lucero
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

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Re: Contamination of Arabidopsis seedlings

Allan-38
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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