Team:Grenoble/Biology/Protocols/Gel

From 2012.igem.org

iGEM Grenoble 2012

Project

Gel electrophoresis

Goal

Separate DNA strands of different lengths.

Prepare the gel

  1. Add 1 g of agarose powder to 50 mL of 1X TAE buffer.

  2. Heat the solution using the microwave until the agarose is completely dissolved.

  3. SAFETY AND USEFUL RECOMMANDATION
    When heating the mixture it is important to use an adaptable container. Putting metallic objects in the microwave is dangerous. It causes the formation of intense electric current. Keep in mind that the container might explode causing the deterioration of the microwave, the preparation and possible injuries to the people next to the device. To avoid this kind of situation the microwave is only used for short heating periods with the user keepin a eye on the preparation.

  4. Once the solution's temperature has decreased (enough to be able to touch the container), pour it into a gel cast (and don't forget the comb).

Load the gel

  1. When the gel is solid, carefully remove the comb.

  2. Place the gel in the buffer tank and cover it with the 1X TAE buffer solution.

  3. Load each sample into the wells (and don't forget the loading dye and the DNA ladder).

Run the gel

  1. Connect the generator.

  2. Chose the right voltage.

  3. Turn on the device.

  4. SAFETY AND USEFUL RECOMMANDATION
    The generator can deliver over 130V. The risk of electrocution for people is generally around 50V. That is why some recommendations must be followed. Before using the electrophoresis device, users should check if they have all parts of the device, because they are removable. They also have to check that each part corresponds to the right device. There are different kinds of tanks and parts are not compatible with each other. Using parts from another type of tank may lead to the loss of safety protections. We managed to assign a number to each corresponding part in order to know to which device they belong to. The generator must not be in presence of water and its vents must not be blocked. The generator has two main safety protections: a detector of migration buffer and a lid. The detection does not let the device generate electricity until the migration buffer has been put in the tank and reached the correct level. This is a protection both for the device and for users. Moreover, it prevents users to be surprised by an unexpected production of electricity during the manipulation. The second protection prevents the generator to produce electricity when no lid is put on the tank. With this, there is less risk of injuries and accidents, if it's correctly used.