Well then...

Well, there may be some issues with my extraction technique. As the protocol I used will be undergoing some drastic changes, and could possibly be scrapped completely for another method, I will give just an overview of what is going on.

The above sample of leaf material that I collected was ground to a slurry in a lysis buffer solution that served to extract the DNA from other plant cell material. DNA is a negatively charged molecule and there are positively charged "beads" in the solution that adhere to them. The entire solution was then centrifuged to separate the beads and DNA from the heavier plant material. After filtration and a wash with isopropanol, I should have had a viable sample of DNA to run gel electrophoresis.

In gel electrophoresis, DNA is loaded into an agarose gel and current is applied (in this case 120 volts for 30 minutes). As I mentioned before DNA holds a negative charge and by placing it in the gel near the negative probe, it is drawn through the gel towards the positive probe. As DNA travels through the gel segments with more base pairs (Adenine with Thymine, and Guanine with Cytosine) will travel much slower than those with fewer base pairs, and the bands can be then observed and recorded.

This is the gel box I used. Apologies for the poor quality picture, I believe I deleted the wrong picture.

In my case, this did not go so well. The DNA sample did travel, but it went the wrong way! THe sample traveled to the negative end of the gel. This indicates to me that the positively charged beads used in the extraction actually made it through the filtration and wash process and were still bound to the DNA and pulled it to the negative pole. After discussion with Anil, and Matt, we have a couple different ways to change our extraction protocol. We think that the protocol we used was designed more for animal cells than plants, and the filtration wasn't up to par with the plant material.

I am not sure when I will continue with the extractions as the weight of the end of the semester is upon us. My research paper on the bacterial identification draft needs some polishing into a final draft, and I still need to develop an oral presentation of it as well (not to mention papers due in my other classes and impending finals). This extended Thanksgiving weekend will not be quite as relaxing as those passed...

HAPPY THANKSGIVING ALL!