Quantitative binding of covalently closed circular DNA to nitrocellulose in NaI.
A procedure for quantitatively binding covalently closed circular DNA to nitrocellulose is detailed. The procedure involves making DNA saturated with respect to Nat at 25 degrees C, denaturing the DNA by heating to 80-100 degrees C, then passing the DNA-NaI solution through a nitrocellulose membrane at 50-60 degrees C. Form I DNA remains denatured throughout the loading procedure and is quantitatively deposited on the membrane as denatured DNA. The mechanism of binding DNA to nitrocellulose in NaI appears to be different from the binding promoted by NaCl.
D003101:Collodion; D004270:DNA, Circular; D007454:Iodides; D008567:Membranes, Artificial; D009691:Nucleic Acid Denaturation; D009693:Nucleic Acid Hybridization; D012974:Sodium Iodide
D004270:DNA, Circular; D007454:Iodides; D008567:Membranes, Artificial; D003101:Collodion; D012974:Sodium Iodide