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 .
