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 the small gel box, you want to use 1.2 gm of the gel powder.

1. After weighing the amount of gel powder required pour in to a beaker containing the required (60 ml/100 ml) of 1x TAE to make a solution. Add the required amount of die (Use the table above).

2. Microwave the solution for about 2 min. You should see a clear solution after boiling it. 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 the rubber washers to make a gel case. Now insert the appropriate comb and pour the clear solution in to the case. A lane from a 1.5 comb can hold 60 ul of sample.

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

5. 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 in to 60 ul lane.

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

7. Run the gel at constant voltage around 175 V for > 20 min.
    Warning: 155V can make the gel hot and warp your bands. If this is concern, use a lower voltage.