Running a sucrose gradient and I am only interested in the disome peak, would also like to separate out the monosome peak. Not interested in polysomes past this.
We usually run a gradient 10-50% sucrose to get the full range of ribosomes - from subunits to the high polysomes.
Should I attenuate the glucose density gradient for this more narrow focus? Would 10-30% be reasonable?
Hey Aditya,
when I was running ribosome profiling from mammalian cells, the 80S monosome and disomy peaks popped up approx. in the middle of 10-50% sucrose gradients (SW40 rotor, 35 000 rpm, 1 h 40 min), so 10-30% sounds like a good starting point to me.
Do you have equipment for fractionation and simultaneous OD measurement?
If yes, it would be easiest to give it a test run to be on the safe side :)
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