Running of agarose gels
Separation of DNA and PCR products was performed by electrophoresis using agarose gels. The density of gel used depends upon product length. DNA and PCR samples are prepared for loading by the addition of 10X loading dye (30% glycerol, 0.25% Xylene Cyanol, 0.25% Bromophenol Blue) and dH 2O.
Constructing agarose gels
To make up the agarose gels for electrophoresis, agarose and 0.5X TBE are mixed to the relevant concentration (typically 0.8 to 4% w/v). The solution is heated in a microwave oven until boiling. The solution is allowed to cool to approximately 65 oC and ethidium bromide added. The solution is poured into a gel holder and allowed to set. At this stage it is placed into a gel tank (Bio-Rad) containing 0.5X TBE buffer. Following loading, the samples are electrophoresed at between 50V and 125V on the agarose gels. Following electrophoreses DNA and PCR products are visualised in UV light by use of a UV transilluminator, and the image captured using the Gel DOC 2000 TM (BIO-RAD) system.