Beginning A Trek Into A New Naboo

During the week of November 1st, I began the protocol for extracting DNA from the cholla cacti. I received a piece of cholla from our green house in hopes of extracting DNA and finding clear polymorphic banding after running the DNA sample in a gel. There were various ways on how I began isolating the DNA.

• The first protocol I used consisted of washing 5-15 spines in a microtube and washing them in bleach and decant with a disposable pipet.
• I rinse the spines with distilled water twice and decant.
• I cut the spines with nail-cutter scissors.
• I added 1000 micro-liters of lysis buffer after the spines are cut and incubate for 30 minutes at 65 degrees Celsius. 
• I disrupt the spines by vortex machine and incubate for another 30 minutes.
• I then incubate the sample overnight by allowing it to sit at room temperature.
• I add 5 drops of protease to the microcentrifuge-tube and heat for 10 minutes at 65 degrees Celsius.
• I vortex the sample and put it in the centrifuge at 13.2 rpm for 4.0 minutes.
• I take the sample out of the centrifuge and remove the supernatant from the top in order to transfer into a new microcentrifuge-tube. 
• I add -20 degrees Celsius isopropyl-alcohol equal to the amount of supernatant and put the tube at -20 degrees Celsius for 30 minutes.
• I centrifuge the sample and collect the supernatant into a new tube.