GSM9635794: SAMPLE 73; Homo sapiens; miRNA-Seq
Frozen tracheal aspirate (TA) samples were thawed on ice, and mixed by pipetting. For each sample, 300 uL of the aspirate was transferred to a 1.5 ml tube and 75 ul of ExoQuick Exosome Precipitation Solution (System biosciences, cat # EXOQ5A-1) was added. Sample and solution were mixed well by inverting, incubated upright and undisturbed at 4 C overnight. Sample mixture was then centrifuged at 1550 x g for 30 min at 4 C. After centrifugation, supernatant was discarded without disturbing the pellet. Pellet was then resuspended in 100 uL 1x PBS. All 100 uL of the sample was used for RNA extraction. RNA was extracted using Zymo Research Quick-RNA Micro Prep Kit following manufacturer's protocol. The typical size distribution for the small RNA ranged between 13.14 and 283.50 ng. Frozen saliva (SA) samples were thawed on ice. RNA was extracted using Quick-RNA Microprep purification kits (Zymo Research). RNA Pico BioAnalyzer (Agilent technologies) was used to determine the RNA quality and quantity. The typical size distribution for the small RNA and quantities for saliva ranged between 33.90 and 922.50 ng Small RNA libraries were prepared from 5–250 ng total RNA using the QIAseq miRNA Library Kit (QIAGEN) as per the manufacturer's instructions.