Cleaning of the Nuclear Fraction (FOCUS™ SubCell Kit)
Resuspend the nuclear pellet in 300µl SubCell Buffer‐III.
Using a sharp pipette tip, remove the sticky lump if any.
The lump is formed from dead cells and some lysed nuclei.
Centrifuge the tube at 700x g for 5 minutes and discard the supernatant.
The pellet containing nuclei is clean enough for most purposes.
If further cleaning required, go to next step.
Pipette 300µl SubCell Buffer‐IV to a 1.5ml centrifuge tube.
Resuspend the nuclear pellet in 100µl SubCell Buffer‐III.
Carefully overlay the nuclear suspension on the surface of SubCell Buffer‐IV.
Centrifuge the tube at 1,000x g for 10 minutes.
Remove the supernatant and collect the very clean nuclear pellet in the tube.
