Week 10. Semester 2. Failed.

After incorporating an incubation period of 1 hr and overnight, respectively, in a solution containing Aspergillus cellulase complex to my previously used protocol, no DNA could be detected under UV light.

Unfortunately, there were a few mishaps in this run. To list a few:

- Hot plate was left on at 65 degrees Celsius for the overnight incubation, but I failed to leave a note and after Mat (the only one aware) left, someone turned it off, which probably was the right thing to do since it had no "please don't turn off" note.

- After the cellulase and lysis buffer incubations, there is a centrifuge step and the supernatant is to be kept because all the cellular debris pellets in the bottom. Accidentally I discarded the supernatant.

- For the overnight extraction, there are two PCI then chloroform extractions both followed by an alcohol addition step. To save time I thought I would synchronize the extractions and ended up unintentionally skipping the second PCI & chloroform extraction.

-The Optimal pH (5.0) and Temp (65C) I used for the cellulase incubation were based on information provided from the Manen et all. 2005 research article I cited on my last post. The source organism was the fungus Trichoderma. While recording my results I found a note I had made to myself about the optimal pH and Temp of Aspergillus. Definitely not the same...


All things considered I kinda knew I wouldn't find anything. For next week I will modify the incubation time and temperature, as well as the cellulase buffer and see what happens. Please take a look at my failed extractions. The one showing DNA is actually my control from a previous extraction.

   Only bottom has visible DNA and is from previous extraction.

I chose to have a Grind and No Grind sample.