Protocol Online logo
Top : Forum Archives: : Molecular Cloning

I added 2.5 times as much LB to TSS (Transformation and Storage Buffer) - What would happen? (Aug/15/2007 )

What happens if I added 2.5 times as much LB to the TSS that I use for making + storing competent cells?

I would discard it if it is not good...

Also, I once accidentally autoclaved LB medium without opening the bottle. The solution I get is much darker than normal. Can I still use it?

Thanks biggrin.gif



I am unfamiliar with chemical comptent cells... but I say the prognosis is not good. Best to discard the medium. If I recall correctly TSS buffer has DMSO and PEG.

Double autoclaving the media will lead to lower cell yeilds. However, depending on the use, it may not matter. (For instance if you are growing small high copy number plasmids, there is no noticible difference. However use said medium for low copy number plasmid, and you will noticibly lower yeilds.)