title: GSM5260225: HCT116-TAK-1; Homo sapiens; RNA-Seq
library_construction_protocol: Cell pellets were collected after centrifuge, flash frozen on dry ice, and total RNA was extracted using RNeasy Mini Kit (Qiagen, 74104). Illumina TruSeq RNA Library Prep Kit (Cat#RS-122-2001) was used with 1 μg of total RNA for the construction of sequencing libraries. RNAseq libraries were prepared for sequencing using standard Illumina protocols. Briefly, first polyA containing mRNA molecules were purified using oligo-dT attached magnetic beads from total RNA and then fragmented. The fragmented mRNA were reverse transcribed into first strand cDNA with reverse transcriptase and random primers. Then the RNA template were removed and replaced by synthesis of the second strand cDNA. Then the end of the cDNA fragments were repaired into blunt ends and adenylated at 3' ends. Then the indexing adapters were ligated to the ends of cDNA fragments, and the DNA fragments with adapters on both ends were enriched and amplified by PCR. After clean-up, the PCR products go through quality control and quantitation to get the final library. Sequencing were run on Novaseq S4 platform: PE150.