Purification of RNA by SDS solubilization and phenol extraction.
This protocol describes a method for RNA purification by sodium dodecyl sulfate (SDS) solubilization and phenol extraction. It is of wide utility and is used routinely to deproteinize RNAs in biological material that has been solubilized in SDS, an ionic detergent that dissolves membranes, disrupts protein-nucleic acid interactions, and inactivates ribonucleases. Once solubilized, addition of phenol or phenol:chloroform:isoamyl alcohol (PCA) completely denatures the protein, and it becomes insoluble in aqueous solution. PCA extraction is the method of choice for preparing cytoplasmic RNA from tissue culture cells or in any other situation (e.g., enzyme reactions) where solubilization in SDS is easily achievable.
Donald C
DC
Rio
Manuel
M
Ares
Gregory J
GJ
Hannon
Timothy W
TW
Nilsen
D000818:Animals; D008967:Molecular Biology / Q000379:methods*; D019800:Phenol / Q000737:chemistry*; D012313:RNA / Q000302:isolation & purification*; D012967:Sodium Dodecyl Sulfate / Q000737:chemistry*; D012995:Solubility
D019800:Phenol; D012967:Sodium Dodecyl Sulfate; D012313:RNA