Ambient Temp, High CO2, 6hrs, rep2
Frozen coral branch fragments were fractured using a flame-sterilized hammer and chisel. To homogenize the samples, fractured coral pieces were placed in 2 mL Eppendorf tubes with 600 micro L Buffer RLT Plus (Qiagen) lysis buffer and ~100 micro L 0.5 mm zirconia/silica beads (BioSpec Products) and vortexed for 5 minutes. Cell debris and coral skeletal fragments were pelleted by centrifugation and the lysate was removed for RNA extraction using the AllPrep DNA/RNA/miRNA Universal Kit (Qiagen). Complementary DNA (cDNA) libraries were prepared from RNA extracts using the TruSeq RNA Sample Prep Kit v2. A total of 21 cDNA libraries were sequenced on an Illumina MiSeq platform using 75 x 75 paired-end cycle kits.