GSM4155757: symsym rep3; Exaiptasia pallida; RNA-Seq
Following collection of cells, these were lysed in 4.3 µl lysis buffer (0.2 % TritonX-100, 1U/µl Protector RNase inhibitor (3335399001, Sigma-Aldrich Co. LLC), 1.25 µM oligo-dT30VN, and 2.5 mM dNTP mix), and flash-frozen in liquid nitrogen. Sequencing libraries were prepared following the SmartSeq2 protocol (Picelli et al., 2014). Briefly, full-length polyA+ mRNA was reverse transcribed using SuperScript II Reverse Transcriptase (18064014, Sigma-Aldrich Co. LLC), followed by pre-amplification of cDNA over 21 PCR cycles using KAPA HiFi Hotstart Ready Mix (07958935001, Roche Diagnostics International AG, Rotkreuz, Switzerland). cDNA libraries were then prepared for Illumina sequencing using the Nextera XT sequencing kit (FC-131-1096, Illumina, Inc., San Diego, CA, USA) and sequenced on NextSeq500 with 75 bp paired-end sequencing.