bacteria in wash buffer

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so i made this wash buffer for western blotting two weeks ago and didnt use it after that. it was still in the 2000 ml cylinder. anyway, today there was some white bacteria floating inside... A LOT of white bacteria... stuck on the cylinder too... does anyone know what kind it is? and how it grew in there?

thanks

 

[vcatz]

No idea what type of bug is in there but your glassware or one of the tools you used to make the buffer (spatula, stir bar, etc...) were probably contaminated. I usually make all Western buffers up fresh and use them right away.

 

[PostalWookie]

Just make a 50x or 10x stock and dilute it fresh each time. If it's that concentrated, nothing should grow in there, and if soemthing does, maybe you should shift your research to study that.

 

[stoic]

Just make a 50x or 10x stock and dilute it fresh each time. If it's that concentrated, nothing should grow in there, and if soemthing does, maybe you should shift your research to study that.

Ditto.

My money for the source of contamination is the graduated cylinder(s). They don't always get autoclaved (at least in my lab).

 

[sunsweet]

I use PBST (or TBST) for washing buffer and its fine even after a long time. so as the others said, it was probably one of the graduated cylinders/instruments you were using.

 

[pratik7]

i would guess that the bug was a halophile of some type

 

[patzan]

it is most likely a fungus, actually. Happens quite often in undergrad biochem labs.

 

[katiekate]

I use TBST w/Tween20. Never had any problems with contamination though. I don't know how anything could grow in there anyway. Wash your container and graduated cylinder and use a cap for the cylinder.