GSM8827685: TRNA70_S50_R1_001; Homo sapiens; miRNA-Seq
Once thawed, the Schirmer strips were removed from the tube using RNase-free pipette tips (Fisher Scientific), placed in a 0.22-µm cellulose-acetate filter tube (Fisher Scientific), and centrifuged at 12,000 x g for 15 minutes at 4 °C. The eluate from the filter tube was collected and returned to the original collection tube containing any leftover volume not absorbed by the Schirmer strip. RNA extraction was performed using the miRNeasy Serum/Plasma Kit (Qiagen) according to the manufacturer's protocol Briefly, the mature miRNAs from 10 ng of total RNA were ligated sequentially with the specially designed 3' and 5' adaptors and converted to cDNA using RT primer integrated with unique molecular indices (UMI) to enable quantification of individual sncRNA molecules. The cDNA products were then purified, enriched by PCR, and purified using QMN Beads (Qiagen) to create the final cDNA library.