A one step method for the recovery of unstained DNA from agarose gels using pBR322 as a mode

Abstract

The isolation of specific fragments of DNA from agarose gels has become a necessity in current molecular biological research. There are, however, several problems associated with the recovery of DNA from the gels, including: 1) contamination of DNA fragments with intercalating dyes and surrounding medium, 2) failure to quantitatively recover high molecular weight DNA, and 3) the need for expensive equipment and time consuming procedures. This study discusses a rapid simple, and efficient technique for recovery of specific unstained and uncontaminated DNA from agarose gels. Electrophoresis is initiated in a modified casting tray, directly on an ultraviolet transilluminator. The progress of unstained bands are monitored via the use of pre-stained DNA in a parallel lane. The DNA is then recovered by direct pipette aspiration from the preformed well with high efficiency. Additional purification, concentration, and manipulation of the recovered DNA is not necessary, and the recovered DNA can be utilized immediately.