Preparative and analytical purification of DNA from agarose.

TLDR

Two NaI‑based procedures were developed: a preparative method that binds DNA to glass for rapid, high‑yield recovery, and an analytical method that precipitates DNA with acetone for quantitative hybridization scanning. The procedures yield high‑yield, agarose‑free DNA that remains enzymatically active, and the acetone‑precipitation method provides a quantitative hybridization assay ideal for restriction digest analysis.

Abstract

Two procedures were developed for removing DNA from agarose after electrophoretic separation of DNA fragments according to size. Both involve dissolving the DNA-containing agarose in NaI. The preparative technique uses binding of DNA to glass in the presence of NaI. The method is rapid and convenient, and DNA of all molecular weight ranges can be recovered in high yield and without degradation. The DNA is free of agarose and remains susceptible to digestion by restriction enzymes. The analytical technique uses selective precipitation of DNA with acetone and has been adapted to molecular hybridization scans of sequences in agarose gels. The sequence-monitoring system is quantitative, directly measuring the proportion of the probe complementary to a given DNA fragment and vice versa. It is especially suitable for analyzing restriction enzyme digests of DNA in mapping experiments.