Binding and eluting a DNA binding column - (Nov/24/2009 )
I have a very general question. When passing a DNA containing sample through a standard DNA binding column, is it better to centrifuge at a slower speed? Obviously it is probably best to just follow the protocol, but I figured if the solution is moving through the column slower, then the DNA has a longer time to bind more to it. Similarly, during the elution step, if I spin the elution buffer through the column slower, then the DNA has more time to interact with it and fall off the column. Does anything I'm saying make sense or are these minor increases in time irrelevant?
you said it: "it's probably best to follow the protocol". the protocol for kits have been worked out, extensively, and have been optimized, in general.
you should only fool around with the conditions if your results are not in accordance with the protocol.