RNA-Seq
The whole venous blood from 2 healthy control, and 6 representative ECMO patients (3 ECMO_S, and 3 ECMO_D) was collected in PAXgene Blood RNA tubes (QIAGEN, Hilden, Germany), and stored at -80C, following the manufacturers instructions. Briefly, the RNA extraction involved thawing samples, lysing cells, and purifying nucleic acids. The RNA yield was estimated by measuring the absorbance at 260nm in a NanoDrop 2000 spectrophotometer (Life Technologies, Carlsbad, CA, USA). RNA purity was calculated from the ratio of absorbance at 260 and 280nm, while RNA integrity was assessed using the 4200 Tape Station System (Agilent Technologies, Palo Alto, CA, USA), which provides RNA integrity number (RIN) scores for RNA quality control. At last, the libraries were generated for sequencing using TruSeq mRNA V2 sample preparation kit with Ribo-Zero Gold (Illumina, San Diego, CA, USA), following the manufacturers instructions. Qubit 2.0 Fluorometer (Life Technologies, Carlsbad, CA, USA) and 4200 Tape Station System (Agilent Technologies, Santa Clara, CA, USA) were used to assess the quality and yield of the prepared libraries. Sequencing was done on a NextSeq 550 (Illumina, San Diego, CA, USA) with 276 cycles, following the manufacturers instructions
SUB15732936
CV27