title: GSM8800666: Healthy control 4; Homo sapiens; RNA-Seq
library_construction_protocol: Total RNAs were extracted fromsaliva samples using the RNeasy kit (Qiagen, Hilden, Germany) using 700 μL saliva from each sample. The extracted RNA was then quantified using a NanoDrop spectrophotometer (ThermoFisher Scientific, USA) . All the extracted RNA was stored at -80 ºC. Genomic DNA removal from RNA samples was ensured by treating the RNA with Turbo DNase (ThermoFisher Scientific, USA). Next, whole transcriptome sequencing was performed using targeted RNA-Seq with Ion AmpliSeq whole transcriptome human gene expression kit (ThermoFisher Scientific, USA). Briefly, cDNA was synthesized using a SuperScript VILO cDNA Synthesis kit (ThermoFisher Scientific, USA) and amplified using Ion AmpliSeq human gene expression core panel primers. The amplified products were subjected to enzymatic shearing to get amplicons of ~200 bp, then ligated with the adapter and the unique barcodes. Next, the constructed library was purified using Agencourt AMPure XP Beads (Beckman Coulter, USA), quantified using an Ion Library TaqMan™ quantitation kit (Applied Biosystems, USA), further diluted to 100 pM, and pooled equally with 16 individual samples. The diluted library was amplified and enriched in Ion Chef System (ThermoFisher Scientific) according to the manufacturer's instructions. Ion 540 Chip was used to sequence the prepared template libraries using an Ion S5 XL Semiconductor sequencer (ThermoFisher Scientific).