Illumina RNA-seq of Pocillopora meandrina transcriptome
RNA was extracted from the coral fragments that had been snap frozen and stored at -80C. A small piece was clipped off using clippers sterilized in 10% bleach, deionized water, isopropanol, and RNAse free water, and then placed in 2 mL microcentrifuge tube containing 0.5mm glass beads (Fisher Scientific Catalog. No 15-340-152) with 1000 microliters of DNA/RNA shield. Tubes were vortexed at high speed for 1 minute before the supernatant was removed. Subsequently, 300 microliters of sample from the homogenate was extracted with the Zymo Quick-DNA/RNA Miniprep Plus Kit Protocol with the following modifications. A first elution of 10 microliters of warmed 10 mM Tris HCl was added to DNA columns for a 15-minute room temperature incubation and centrifuged at 16,000 rcf(g) for 30 seconds. A second elution of 100 microliters of warmed 10 mM Tris HCl was added for a 5-minute incubation and centrifuged at 16,000 rcf(g) for 30 seconds. RNA quantity (ng_microliter) was measured with a ThermoFisher Qubit Fluorometer high-sensitivity kit. RNA quality was measured with an Agilent TapeStation System. DNA was send to DNA Link Sequencing Lab (https://www.dnalinkseqlab.com) for sequencing on their Illumina NovaSeq 6000 platform.