New Protocol Redux

Last week I spoke about being very happy with a new extraction protocol that I had found. Scratch that. Upon further investigation into it, I relised that the protocol listed all of the chemicals required for each of the buffers used, but failed to itemize the ratios of these chemicals in the solutions. Essentialy, I had the ingredients without the recipe.

After several more hours of digging I came upon a great resource website: http://www.protocol-online.org. Through this site, I came upon an outstanding list of DNA extraction protocols from the University of Wisconsin-Madison: http://labs.medmicro.wisc.edu/mcfall-ngai/papers/2002nish3.pdf. From this list I will be using protocol 17, sans the liquid nitrogen. The "outstanding" part of this list is the detailed list of exactly how to make many of the common reagents for DNA extraction. The shopping list begins.

Last Friday I did make it back out to my sampling site at Squaw Peak and collected fresh specimen from the same subjects I have been working with. I was more than a bit worried that the recent monsoon activity would negatively impact my field location as it is a wash. At first blush, there did not seem to be any major changes, but I was unable to observe the site closely as I was pressed for time. I will be returning in the not to distant future to more closely examine the site for changes (are more of the root systems exposed showing the relation between individuals). I spent a little over an hour collecting leaf samples, washing them in ethanol, and labeling them. They are currently in storage at -80 degrees Celsius.