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mRNA Isolation Protocol
A. Reagents:
For mRNA isolation
Binding Buffer OBB
20 mM Tris-Cl pH7.5, 1 M NaCl, 2 mM EDTA, 0.2% SDS
Oligotex Suspension
10% (w/v) in 10 mM Tris-Cl pH7.5, 500 mM NaCl, 1 mM EDTA, 0.1% SDS, 0.1% NaN3.
Wash Buffer OW2
10 mM Tris-Cl pH7.5, 150 mM NaCl, 1 mM EDTA
Elution Buffer OEB
5 mM Tris-Cl pH 7.5
Small spin columns
RNase-free spin columns
For mRNA precipitation
4M LiCl
Linear acrylamide (5 mg/ml)
100% EtOH
Oligotex mRNA kits Mini (70022, <250µg), Midi (70042, 250µg-1mg) and Maxi (70061, 1-3mg)