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-P/C/I is the optimal choice for RNA isolation and other procedures where a low phenol pH value is desired.
The low pH results in interphase enrichment of the DNA, reducing DNA contamination in the RNA eluate.
Addition of 1 volume ROTI®Aqua-P/C/I and thorough mixing. Phenol and chloroform lead to denaturation of proteins which accumulate in the interphase. Isoamyl alcohol prevents foaming and very efficiently inhibits RNAses. 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-P/C/I 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.
Bottled under argon. |