Routine isolation of the 70 S RNA of a murine oncornavirus by use of an unesterified cellulose column

Chemical linkage of nucleic acids to neutral and phosphorylated cellulose powders and isolation of specific sequences by affinity chromatography

“…The DNA or RNA immobilized on cellulose is apparently free to interact with complementary sequences in the mobile phase; a gel matrix imposes an additional permeability barrier. Although denatured DNA or mRNA containing poly(A) stretches exhibits strong affinity for cellulose preparations with ligninlike impurities or possessing a microcrystalline structure (Kitos et al, 1972;DeLarco and Guroff, 1973;Larsen et al, 1973;Sullivan and Roberts, 1973), nonspecific binding can be reduced by using highly purified fibrous cellulose powder and extensively washing of the column with a low ionic strength solution {e.g., 0.2 or 0.1 X SSC) at 4°p rior to the elution of the retained hybrids. Due to the charge repulsion at low ionic strength, the additional phosphate anions present in phosphocellulose may help in reducing the nonspecific adsorption of polyanionic DNA or RNA to the solid supporting medium.…”