During the week of November 12, I continued trying to perfect my Cacti DNA extraction. This new protocol involved my older protocol to extract DNA from ants. This protocol involved:
- I put 5-15 spines into a microtube and wash with bleach.
- I then decant and rinse the spines twice with distilled water.
- I cut the spines into smaller pieces with nail-cutter scissors.
- I add 90 micro-liters of nuclease free water, 1 micro-liter of 10% SDS, 2 micro-liters of 20% meat tenderizer solution.
- I heated the tube at 75 degrees Celsius for 10 minutes in a water bath and agitated the tube thereafter with a flick of my finger.
- I heated the tube at 95 degree Celsius for 5 minutes in a heating block.
- I agitate the tube with a flick of my finger then allow the tube to incubate in room temperature overnight.
- I take the sample and put it into the centrifuge for 13.2 rpm for 5 min.
- 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.
The heating block used and the other picture shows two samples with -20 degree Celsius isopropyl-alcohol.
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