Cleaning of the Nuclear Fraction (FOCUS™ SubCell Kit)
Resuspend the nuclear pellet in 300µl SubCell Buffer‐III.
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.
Using a sharp pipette tip, remove the sticky lump if any.
The lump is formed from dead cells and some lysed nuclei.
Remove the supernatant and collect the very clean nuclear pellet in the tube.
