TSS Transformation troubleshooting
Posted 19 January 2011 - 05:34 AM
Posted 19 January 2011 - 05:57 AM
Posted 19 January 2011 - 06:05 AM
Usually even bad transformation yields a few colonies. Make sure your plasmid antibiotic resistance and the plate antibiotic agree. Are you recovering the cells for an hour on non-selective medium? Can you transform other cells with this plasmid and the same plates?
yes .i am growing them on SOC for one hour.My ligations are ok.antibiotics are ok.
Posted 19 January 2011 - 02:14 PM
Posted 19 January 2011 - 07:10 PM
So, are you trying to transform ligation products, or purified plasmid? If you are having trouble, I would highly suggest transforming serial dilutions of a known working plasmid, rather than starting with ligation products. Too many things can go wrong with ligations, and you want to separate the source of the problem. You should be able to easily transform 100 pg of purified plasmid DNA to get hundreds of colonies.
I will try with this .thanks for suggestion.