How to dissolve over-dried DNA?
Posted 28 March 2005 - 02:01 PM
Posted 28 March 2005 - 10:14 PM
what have you dissolved the pellet in?
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Posted 29 March 2005 - 01:42 AM
I guess you dissolved your plasmid pellet in ddH2O, right?
Try to dissolve in 8mM NaOH.
Next time when you extract any of your DNA, never never let it over dry.
After you discard the supernatant (70% ethanol) try to pipet out the what ever solution remain at the bottum of the tube and let it air dry brifly. Or you can put on thermal cycler block, set at 50-60oC for 2 min (I always do so).
Posted 29 March 2005 - 02:13 AM
Posted 29 March 2005 - 02:28 AM
hum when nucleic acid forms an unsoluble pellet (lokks like a gel?) it's probably RNA. If you want to check it, try to dissolve it in a small amount of water (150Ál ddH2O) and heat it to 60░ duting 10'. If dissolving well, check on agarose gel.
Posted 29 March 2005 - 03:01 AM
when there is lot of DNA it may take time to dissolve. I usually leave it at 4 C overnight. Better not to disturb it by vortexing and trying to brake the clumps etc. if it is DNA should has to dissolve, isnn't it?
Posted 29 March 2005 - 12:41 PM