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Phenolic DNA Isolation
Background and protocol
Phenol and chloroform lead to denaturation of proteins which accumulate in the interphase. Isoamyl alcohol prevents foaming and very efficiently inhibits RNAses.
For distinct phase separation, ROTI®Aqua-Phenol needs a pH of ≤4.0 in the aqueous upper phase. If needed, the upper phase may be acidified by adding 1-2 drops of 1 N HCl.
Phenol solutions for nucleic acid isolation must be buffered and adjusted to the correct pH value prior to use. ROTI®Phenol is a redistilled, in TE buffer equilibrated readytouse phenol.
ROTI®Phenol has been successfully tried and tested for isolating nucleic acids in many research laboratories. This quality product for molecular biology replaces the time consuming preparation of phenol and subsequently any exposure to toxic vapours.
Draw solution from lower phase. Do not shake before use!
Addition of 1 volume ROTI®Phenol and thorough mixing. Phenol leads to denaturation of proteins which accumulate in the interphase. Following centrifugation, the nucleic acid containing solution may be retrieved as upper phase.
Bottled under argon. |