Please note our Technical Information Brochure on the Internet next to the products:
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.
ROTI®Aqua Phenol is perfectly suited for RNA isolation and other procedures, requiring a low phenol pH value. The low pH results in interphase enrichment of the DNA, reducing DNA contamination in the RNA eluate.
Also ideal for self-adjustment of the pH-value. If required, buffer solution (e.g. TE buffer) may be added directly into the bottle, as ROTI®Aqua-Phenol comes in the following bottle sizes:
A980.2: 100 ml in a 250 ml bottle
A980.1: 250 ml in a 500 ml bottle
A980.3: 500 ml in a 1 l bottle
Addition of 1 volume ROTI®Aqua-Phenol and thorough mixing. Phenol leads to denaturation of proteins which accumulate in the interphase. The acid pH-value causes a depletion of the DNA, so that only little contaminating DNA is isolated. Following centrifugation, the RNA containing solution may be retrieved as upper phase.
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 12 drops of 1 N HCl.
Draw solution from lower phase. Do not shake before use!
Bottled under argon. |