I have been making NGS and WGBS Libraries for a while now and keep noticing an odd trend with my SPRI beads at the Indexing PCR step. When I go to elute my final Library off the SPRI beads, the beads are harder to resuspend and stiff. They stick to my tip and the tube. I know what you might say: "they are probably over dried," but I do not even air-dry them at this point. I just do a quick spin and remove residual ethanol and quickly resuspend in Low EDTA TE. I was wondering if anyone could give me any guidance on what might be going on and if this is a bad thing that I should be worried about? Is it that the beads are overload with DNA (I doubt this because my library concentrations are on the lower side)? Any guidance on this would be greatly appreciated!
How long do you incubate your library with the beads? What is the volume of ethanol you use for washing and how many times do you wash? Do you re suspend the beads well before adding? Is the beads in date? Why do you not air dry for 30sec?
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