I'm digesting a plasmid for assembly. In my experience, I always have low yields when gel extracting and purifying. Since I am only making one cut in my vector, could I just purify with a column? Any advice would be greatly appreciated, thanks!.
Yes, this is perfectly possible. There are a few small caveats though:
- do not overload the column with too much DNA, as this will result in lower yields
- also do not load an amount of DNA which is too small, this may result in loss of your sample. In my experience it worked better cleaning small samples from a gel.
- also carryover of salts or Ethanol can cause problems
- look up the exclusion size of the column from the manual. They can typically on purify DNA above a certain size and sometimes only work efficiently up to a certain DNA size
- they are quick and convenient, but more expensive than a gel