chromatography:

 

we have to collect the fractions from the column. i wonder what is the difference between;

 

flowthrough fraction, wash fraction and eluate fraction?

 

i think i know what it means with eluate fraction, but what about the other two?

 

hope for replies!

thanks a bunch!

flowthrough= the liquid that went out of your column while the substance of interest stayed bound to the column when you applied the starting liquid mixture

 

normally after binding you wash a column to get rid of immurities

 

wash fraction= the liquid that came out of the column as you were washing it.

 

after washing you apply a elution buffer

 

eluding fraction= what liquid came off the column as a result of the eluding buffer. this should have you substance of interest.

 

You are probably collecting the different fractions to see if you lost alot of protein/dna/substance when you washed by testing wash fraction and to see if the column worked at all by testing flowthrough fraction.

thanks very much for your sharing, Frostrunner!