this is my first time and ny first post in this forum.
I would appreciate if someone could help me. I need to know what is the role of every reagent used in the classical isothiocyanateâ€“phenol:chloroform extraction protocol used in RNA purification.
Here are the reagents:
4 M guanidine isothiocyanate (GITC)
0.02 M sodium citrate
2 M Sodium acetate (pH 4.0)
Phenol (equilibrated to pH 5.3â€“5.7 with 0.1 M succinic acid)
β-Mercaptoethanol (β-ME) (14.4 M)
Glycogen (2 mg/ml)
75% (v/v) ethanol (prepared using DEPC-treated water)
All the best and thanks a lot to everybody.