Agarose Gel preparation

1 gm of a solute in 100 ml of a solvent makes a 1% (w/v) solution. Hence, if you are pouring a 2% gel in to the small gel box, you want to use 1.2 gm of the gel powder.

1. Weigh the amount of gel powder required and pour it in to a beaker containing (60 ml/100 ml) of 1x TAE. Add the required amount of die (Use the table above).

2. Microwave the solution for about 2 min. You should see a clear solution as it boils. If not, shake the beaker and microwave for an additional 30 sec. Be careful while you shake the beaker as the solution can rise up the brim.

3. Use rubber washers to make a gel case. Insert the appropriate comb and pour the clear, warm solution in to the case. Do not let any solution remain in the glass beaker because it solidifies fast and makes it difficult to clean.

4. Let the box aside (or place in the refrigerator) to cool for about an hour or so until it turns in to an opaque gel.

5. After the gel solidifies, fill the gel box with TAE solution until the gel is submerged.

6. While loading the gel, remember that the loading buffer is 6x. Hence, you want to mix 10 ul for a 50 ul sample to load on to a 60 ul well (A well from a 1.5 comb can hold 60 ul of sample).

7. For the 2 log ladder, use 10 ul of the NEB ladder.

8. Run the gel at constant voltage around 175 V for > 20 min.
    Warning: 175V can make the gel hot and warp your bands. If this is concern, use a lower voltage.