Salivary_Gland_Tissue_RNAseq

The minor salivary gland samples of each group were mechanically homogenized using a motorized homogenizer (Thermo Fisher Scientific Inc., Germany) followed by RNA extraction using TRIzol Reagent (Qiagen). RNA was quantified and characterized using an Agilent 2100 Bioanalyzer (Agilent Technologies, Palo Alto, CA, USA) and a NanoDrop (Thermo Fisher Scientific Inc.) Gel electrophoresis (1% agarose formaldehyde) was used as an RNA integrity test. For library preparation, one microgram of total RNA with a RIN (RNA integrity number) value above seven was used. The ribosomal RNA-depleted RNA was purified, fragmented, and reverse-transcribed. RNA-seq libraries were sequenced using the Illumina HiSeq instrument (Illumina, CA, USA) with a 2x 150 paired-end (PE) configuration. Image analysis and base calling were conducted in HiSeq Control Software + OLB + GAPipeline-1.6 (Illumina) on the HiSeq instrument. The sequences were processed and analyzed by GENEWIZ (Suzhou, China). After reads were trimmed and technical sequences (adapters, PCR primers, or fragments thereof, and low-quality reads) were removed, and pass filter data of fastq format were processed by Cutadapt (V1.9.1) to be high-quality clean data