GSM3244543: miRNA-E006AA-hT-ADT-1; Homo sapiens; miRNA-Seq
Total RNA was extracted using Qiagen RNeasy Plus Mini Kit (Qiagen). RNA samples were quantified using Qubit 2.0 Fluorometer (Life Technologies, Carlsbad, CA, USA) and RNA integrity were checked with Agilent 2100 Bioanalyzer (Agilent Technologies, Palo Alto, CA, USA). Small RNA sequencing library is prepared by using Illumina TruSeq Small RNA library Prep Kit (Illumina, San Diego, CA). In brief, Illumina 3' and 5' adapter is added to RNA molecules with a 5'-phosphate and a 3'-hydroxyl group sequentially. A reverse transcription reaction were used to create single stranded cDNA. The cDNA was then PCR amplified using a common primer and a primer containing index sequence. The sequencing libraries were multiplexed and clustered on one lane of a flowcell. After clustering, the flowcell will be loaded on the Illumina HiSeq instrument according to manufacturer's instructions. The samples will be sequenced using a 1x50 Single End (SE) configuration. Image analysis and base calling were conducted by the HiSeq Control Software (HCS) on the HiSeq instrument. Raw sequence data (.bcl files) generated from Illumina HiSeq will be converted into fastq files and de-multiplexed using Illumina bsl2fastq v. 2.17 program. One mis-match will be allowed for index sequence identification.
GEO Accession
GSM3244543