This week I extracted DNA from Porites astreoides samples collected the week of 11/8/16 in Belize. These samples represent the endpoint of a year long common garden experiment in the backreef at Carrie Bow Cay. Detailed sample info can be found in the initial post here. The samples were stored at room temperature in salt saturated DMSO. Tissue was scraped off the skeletons with forceps into 1.5 ml tubes. Tissue was washed three times with PBS via centrifugation before beginning the Qiagen DNeasy extraction method. An overnight proteinase K lysis period at 56ºC was used. DNA was eluted via two passes with 50 µl AE buffer (100 µl total volume). To further clean the DNA, samples were subject to ethanol precipitation using this protocol. Samples were re-eluted in 50 µl AE buffer.
To assess DNA quantity and quality, samples were tested with the Qubit BR DNA assay followed by electrophoresis of the majority of them on a 1% agarose gel with 1X TBE, 135 volts for 30 min.