GSM7843841: unC-185-03, methylseq; Acropora palmata; Bisulfite-Seq
Genomic DNA (gDNA) was extracted from 32 A. palmata fragments (n= 8 per group condition; 16 fragments per genet). We used the DNeasy Blood and Tissue Kit (Qiagen, Switzerland), as per the manufacturer's protocol. gDNA concentration was quantified at 0.5-1.5 μg (Qubit® dsDNA BR Assay Kit on a Qubit® 2.0. Fluorometer) and sent for Whole Genome Bisulfite Sequencing (WGBS) at Admera Health (New Jersey, USA). Sequencing libraries were prepared with an average sequencing library insert size of 450 bp and according to TruSeq® DNA Methylation Kit protocol, Sample Preparation Guide (Illumina Inc., USA). Briefly, standard reaction mix consisting of 130 µl of the CT Conversion Reagent and 20 µl of each DNA sample were used for bisulfite conversion (EZ DNA Methylation-Gold™ Kit) in a thermal cycler (Eppendorf® Mastercycler® Pro S). After the incubation period, bisulfite converted DNA was purified following the protocol of EZ DNA Methylation-Gold™ Kit. The bisulfite converted sequencing library was enriched following the TruSeq® DNA Methylation Kit protocol. Libraries were validated and quantified with qPCR. Sequencing was performed in Illumina Hi-Seq platform with pair-end reads of 2x150 and a mean depth coverage of 30x (Illumina Inc., USA). The estimated number of passed filter reads per sample was 110-120 million paired-end reads (55-60M reads in each direction).