Illumina HiSeq 4000 paired end sequencing; RNA-sequencing of human salivary gland from Sjogren's syndrome and sicca patients
RNA-sequencing of human salivary gland from Sjogren's syndrome and sicca patients
All patients provided written informed consent before the lip biopsy procedure. The biopsies were performed under local anesthesia with a vertical incision on the inner surface of the lower lip.  Following randomization to Rituximab or placebo, patients received two 1g cycles of Rituximab/placebo at baseline and week 24. RNA was extracted using a column-based approach, RNeasy Micro Kit (Qiagen) following the manufacturer's instructions. The RNA samples were quantified using Qubit 2.0 Fluorometer (Invitrogen) and RNA integrity was checked with Agilent TapeStation (Agilent Technologies).  RNA sequencing libraries were prepared using the NEBNext Ultra RNA Library Prep Kit for Illumina. Briefly, mRNA was first enriched with Oligod(T) beads. Enriched mRNAs were fragmented for 15 minutes at 94 °C. First strand and second strand cDNA were subsequently synthesized. cDNA fragments were end repaired and adenylated at 3'ends, and universal adapters were ligated to cDNA fragments, followed by index addition and library enrichment by PCR with limited cycles.
Experimental Factor: disease
Sjogren's syndrome