GSM5768772: L2; Bos taurus; RNA-Seq
Total RNA was isolated from eight pools (four control and four lycopene in four replicates) of ten blastocysts per pool using the RNeasy Micro kit (Qiagen, Germantown, USA) according to the manufacturer's protocol. The quality and concentration of the RNA samples were examined using an RNA 6000 Pico Chip (Agilent Technologies, Carlsbad, CA, United States) and a Quant-iT RiboGreen RNA assay kit (Life Technologies, Carlsbad, CA, United States), respectively. Transcriptome library preparation was done by Qiaseq UPX 3' transcriptome kit (Qiagen) according to the manufacturer's instructions with 10 PCR cycles. Quality of the library preparation was checked with a high sensitivity DNA chip (Agilent technologies) and library quantification was performed by qPCR according to the Illumina qPCR quantification protocol (NXTGNT sequencing facility, Ghent, Belgium), followed by equimolar pooling of libraries based on qPCR. Sequencing was performed on a high throughput Illumina NextSeq 500 flow cell with 20% PhiX spiked in (Read 1: 57 cycles; read 2: 27 cycles and index 6 cycles).