RNA isolation protocol (1)

RNA isolation by the rapid Guanidinium method
Materials
Mononuclear cell pellet isolated from 1 to 2 ml bone marrow or 5 ml whole blood
HBSS without Ca or Mg
RNA isolation solution I (25 g guanidinium thiocyanate (4 M final), 29.3 ml H2O, 1.76 ml 0.75 M sodium citrate, pH7.0 (25 mM final) , 2.64 ml 10% (w/v) N-lauroylsarcosine (0.5% final) dissolve at 65 degree celcius, store at room temperature (stable > or equal 3 months)
Do not inhale quanidinium thiocyanate and wear gloves when handling it, Do not expose guanidinium thiocyanate to acid conditions, because this may produce cyanide gas.
2 M sodium acetate, pH 4
Buffered phenol
24:1 (v/v) chloroform/isoamyl alcohol
100% and 70% (v/v) ethanol
RNA isolation solution II (10 ml RNA isolation solution I, 72 microlitre 2-mercaptoethanol (0.1 M final) Make fresh before each extraction)
DEPC-treated H2O