Help with reagents used in RNA estraction procedure

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Joined: Sun Nov 11, 2007 11:09 pm

Help with reagents used in RNA estraction procedure

Post by Nice2U » Mon Nov 12, 2007 7:20 am

Dear friends,

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 or the function of the reagents used in the classical isothiocyanate–phenol:chloroform extraction protocol used in RNA purification.
Here are the reagents:

4 M guanidine isothiocyanate (GITC)
0.02 M sodium citrate
0.5% sarcosyl
2 M Sodium acetate (pH 4.0)
Chloroform:isoamyl alcohol
Phenol (equilibrated to pH 5.3–5.7 with 0.1 M succinic acid)
β-Mercaptoethanol (β-ME) (14.4 M)
Glycogen (2 mg/ml)
Diethylpyrocarbonate (DEPC)
DEPC-treated water
75% (v/v) ethanol (prepared using DEPC-treated water)

All the best and thanks a lot to everybody.

Posts: 12
Joined: Thu Nov 08, 2007 10:35 pm

Post by benedictine » Mon Nov 12, 2007 11:32 am

hi, actually if u get the Sambrook & Maniatis lab manuals, now I cant remember which volume, comes in 3 volumes, u read that and can find answer there

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