Monthly Archives: July 2015

P. astreoides RNA extraction

Yesterday 7/13/15 I extracted RNA from the two Porites astreoides samples I collected in Belize (see previous post Belize pilot experiments). I used RNAzol RT and followed the total RNA protocol. The 25 mm coral cores were removed from their tubes of RNAlater and placed in a porcelain mortar that had been chilled in the -80C freezer for about 20-30 min before starting (note: this worked well, but the mortar and pestle were not chilled by the time I homogenized the second sample). I scraped off the top layer of tissue and skeleton using a razor blade, removed the rest of the core, and ground the tissue and skeleton in 1 ml RNAzol with a pestle. I did not do the optional phase separation step (#4). I left the RNA pellets in 75% ethanol in the refrigerator. The remaining homogenate in RNAzol was placed in the -80C freezer.