Rapid Non-Enzymatic DNA Deproteinization in Agarose Miniplugs

Abstract

Mini agarose plugs, containing immobilized cells, were lysed and deproteinized in 3–6 h by a non-enzymatic procedure, using 4 M urea and detergents. The performance of Saccharomyces cerevisiae miniCHEF DNA pattern was not affected by the treatment. DNA contained in the miniplugs was completely digested by restriction endonucleases in 1–2 h. This procedure saves time in comparison to the 40 h of incubation with urea used in the original procedure. It also saves reagents and permits a rapid DNA digestion without using PMSF.

Keywords:

• Pulsed Field Minigel Electrophoresis
• Miniplugs, MiniCHEF
• Urea Deproteinization