Week 6

DNA Concentration and Separation by Electrophoresis

At the end of last week, I ran the samples through a spectrophotometer which analyzed the concentration of DNA, RNA, and proteins.This week, I separated these compounds by the electrophoresis method. Bellow is a picture of what the separation of these compounds looked like on the stationary gel used.

One very important thing that I learned this week was to make my very own gel to run the electrophoresis apparatus. I had experience with the electrophoresis apparatus in the past and so i was familiar with the different part of the device and I had an idea of the mechanisms behind this method, that was not new to me.

What was new to me however, was making my own gel. Before I thought that these gels where already fabricated and I was expecting to simply peel one out of some plastic wrapper as if it were a Popsicle. Then, Jasmine Patricio guided me to collect the materials and chemicals necessary for to produce this gel. It was as if we were about to cook a dish, in a sense we were cooking because at one point we heated a mixture containing a solution of TAE and Agarose in the microwave.

The first gel that was produced was not useful because it was too thick and it was missing CyberGreen, a component that stains DNA and it allows it to glow under UV light. That day wasn't very productive as it was near 5 o'clock and we were going to have to home soon.

The next time that I was in lab I prepared the gel successfully on my second try that day.

After getting a gel that would work I ran the separation method with the electrophoresis apparatus. Bellow is a picture of what that run looked like.
The well to the far left was loaded with: loading dye and a molecular-weight size marker also known as a ladder.
The two subsequent where left alone.
The the 4th, 5th, 6th, and 7th well form left to right were filled with my 1st, 2nd, 3rd, and 4th sample of DNA extraction respectively and loading dye.

Something important to note about this result is the size of the ladder under the first well. It looks very bright and short in size, which means it will not be very helpful in determining the size of the bands. One thing that could be improved next time is allowing the eletrophoresis process to run for a bit longer. In this case it was set for 30 minutes, in the futer I think I can double to amount of time and see if that will give for a better latter.