GSM4159298: Colorectal cancer tissue 1; Homo sapiens; RNA-Seq
Total RNA was extracted from tissues using Trizol reagent (Life Technologies, USA). The integrity and quantity of RNA were checked before the sequencing experiment. Due to the decoration of tRNA-derived small RNAs which interferes with small RNA-seq library construction, RNA Pretreatment Kit (#: AS-FS-005, Arraystar, USA) included removing 3'- aminoacyl, 3'-cP, phosphorylates 5'-OH, and demethylates m1A, m1G, and m3C were used in order to get efficient cDNA reverse transcription. After sequentially ligated to 3' and 5' small RNA adapters (#: AS-FS-003, Arraystar, USA), cDNA was synthesized and amplified using Illumina's proprietary RT primers and amplification primers. Subsequently, ~135-160 bp PCR amplified fragments were extracted and purified from the PAGE gel. Finally, the completed libraries were quantified by Agilent 2100 Bioanalyzer (Agilent, USA). The libraries were denatured and diluted to a loading volume of 1.3ml and loading a concentration of 1.8pM. Diluted libraries were loaded onto reagent cartridge and forwarded to sequencing run on Illumina NextSeq 500 system using NextSeq 500/550 V2 kit (#FC-404-2005, Illumina), according to the manufacturer's instructions.
GEO Accession
GSM4159298