GSM3356458: NG-10690_Axin6Ctr; Homo sapiens; RNA-Seq
Total RNA was isolated from 70-80% confluent plates with the Machery-NucleoSpin RNA II kit Library was constructed and sequenced with an Illumina HiSeqTM2000 (GATC Biotech, Konstanz, Germany). Briefly, the mRNA was enriched using oligo-dT magnetic beads, followed by fragmentation (about 200 bp). Then first strand cDNA was synthesized using random hexamer-primers and the second strand was further synthesized in a reaction buffer including dNTPs, RNase H and DNA polymerase I. Double stranded cDNA was purified with magnetic beads. Then, the 3'-end single nucleotide A (adenine) was added and adapters were ligated to the fragments which were enriched by PCR amplification.