Massive Analysis of cDNA Ends of IL-37tg
MACE quantitatively analyzes gene expression based on high-throughput sequencing of a part of the 3’-end of mRNA transcripts. Each sequence-read represents one single transcript molecule. MACE libraries were generated by GenXPro GmbH (www.genxpro.de, Frankfurt, Germany) using the “MACE kit”. Briefly, poly-adenylated mRNA was transcribed into cDNA using biotinylated oligo-dT primers. The resulting biotinylated cDNA was randomly fragmented and the 3’-ends were captured using a streptavidin resin, while the remaining fragments were discarded. A modified “True Quant” Illumina p5 adapter was then ligated to the unbound end of each fragmented cDNA. These constructs were amplified by PCR and sequenced on an Illumina Hiseq 2000 (Illumina), thus generating one single read for each original transcript molecule.