Team:Macquarie Australia/Protocols/Gels


Pre-Stained Gels


Running gels is a time consuming process- in part due to the staining process. For this reason, the gels cast were pre-stained with GelRed. Upon completion of the electrophoresis, the gel can be analysed. Assuming a homogenous distribution of GelRed, the reduction in waiting time makes this methods of casting gels more attractive than post-staining.


A Sub-Cell GT Agarose Gel Electrophoresis system was set up according to the manufacturer’s instructions. The tray used was 15 x 10 cm and required 120ml gel volume to achieve a 1cm thick gel size.

The 1.2% agarose gel was made up to 200ml by combining 2.4g of agarose gel powder with 200ml of previously prepared TAE buffer, GelRed was added to give a final concentration of 1%. This was placed in the microwave to fully dissolve the agar powder. After the gel liquid cooled to around 60°C it was poured into the UV Transparent Plastic (UVTP) tray.

After the gel solidified the system was submerged in TAE buffer. The samples were loaded in appropriate wells and the system was allowed to run for approximately 1 hour, or until bands were well resolved. Following this the gel was removed. The stained gel was finally analysed using a Gel-Doc EZ Imager for visualisation digestion or ligation products.