Nuclear Extraction

-Label 4 tubes per sample, 2 as Cytoplasmic Extract (CE) and 2 as Nuclear Extract (NE)
-Chill tubes in freezer before use
-Trypsinize cells, spin down at 1.5k x g, aspirate trypsin
-Keep cells on ice for the full process
-Add 100 uL CE buffer w/ NP40
-Incubate 3 min on ice
-Spin 4 min at 1.5k rpm at 4 oC
-Remove CE to clean tube
-Gently wash nuclei in 100 uL CE buffer w/out NP40
-Spin as above
-Add 50 uL NE buffer
-Add 35 uL 5 M NaCl
-Add 50 uL NE buffer
-Incubate 10 min on ice, vortexing periodically to resuspend nuclei
-Sonicate NE
-Spin both CE and NE for 10 min at 21.1k g
-Transfer to fresh tubes
-Add glycerol to CE tubes to 20% (~22 uL)
-Freeze