What is Agar?
Agar comes from sea weed. The
gel is 1% agarous and
has no electrical charge.
Why the band get distorted?
The gel mix was clearly incorrect. The acrylamide concentration was so high that only the smallest proteins entered the gel to any extent. The bands were distorted due to overloading and the fact that beyond a reasonable concentration the most concentrated gels tend not to polymerize evenly.
What is significant about the bubbles?
- They indicate that electrolysis of water is taking place.
- One electrode will have a lot of bubbles and the other will have a lesser amount. Why the difference?
- The formula for water is H2O and the splitting of the molecule will produce twice as many atoms of hydrogen.
Why the running process took a longer time than it should?
- Buffers too concentrated. Check buffer protocol; dilute buffer if necessary.
- Current too low. Increase voltage by 25-50%.
What are the causes of frowning effect?
- Center of the gel running hotter than either ends. Decrease power setting. Check buffer protocol to ensure it is properly formulated.
- Insufficient buffer volume. Make sure the gel wells are completely covered with buffer. Check for leaks in the upper buffer chamber that might have occured during the run.
- Electrophoresis buffer was too warm. Use cold electrophoresis buffer.
- Poor quality gel. Bubbles and particles in the gel could cause the smiling effect. Mix and pour all gel preparation solutions carefully to avoid formation of bubbles. If particles are visible in solutions, remove them by filtration.