Quantitative isolation of DNA restriction fragments from low-melting agarose by Elutip-d affinity chromatography.
The use of a disposable affinity column and low-melting-temperature agarose for the quantitative preparation of DNA restriction fragments is presented. After electrophoretic separation of DNA, the band(s) are excised and the DNA/agarose melted in a low-salt buffer. After cooling, the DNA is bound to an Elutip-d affinity column. Fragments are eluted at high salt and concentrated by ethanol precipitation. Recoveries greater than 80% are achieved with purity suitable for most applications in molecular biology.
D002846:Chromatography, Affinity / Q000379:methods*; D004247:DNA / Q000302:isolation & purification*; D004262:DNA Restriction Enzymes; D012685:Sepharose; D013696:Temperature
D004247:DNA; D012685:Sepharose; D004262:DNA Restriction Enzymes