Week 12. Semester 2. Some progress and FUNGUS!!

This week I reattempted a modified version of my protocol utilizing cellulase for breakage of the plant cell wall. The first try was not successful at all due to critical errors during execution and failure to interpret information resulting in an inadequate protocol, using a buffer I failed to dilute probably denaturing the cellulase complex immediately. This week, although not perfect, I finally feel I have made some progress.
The Enzyme incubation period was 24hrs. and I ran both a Grind and No Grind DNA extractions. Another variable was the use of Palo Verde blossoms in place of the leaflets to compare the yields and purity of DNA.
The protocol ran smoothly and I plan on running a gel next Monday to see results. However I can already predict based on the size of the DNA pellet that the samples that were ground in the mortar and pestle has a significant higher amount of DNA. This is great and may prove the enzyme digestion to be a good addition to the protocol. However, my goal is to eliminate the need for grinding altogether, so may be next time I will leave them a day longer. Or I may grind after the cellulase incubation period instead of before?
For next week, I also plant to finally start my fungal culture broths to isolate exoenzymes from them. I have successfully isolated four species from the plates on the pictures below, so we will see. Also Matt brought me samples from a Palo infected with some fungus!!! So I will isolate that as well and go from there hoping it will brake it down. By the way thank you Bethany!! for sharing your beautiful pink plates with me!!!
As we approach the end of the semester, I am feeling very overwhelmed with everything I am involved in at school, my classes and my family. It's hard to find time to sleep and the last couple of weeks were specially difficult since I felt I was not progressing in my project. However, talks with different scholars and with Matt and Josh have lifted my spirits and I feel re-energized and ready to wrap up the semester! I look forward to next semester, when I plan on having additional time to spend in my project!