title: GSM8806078: exo_3b; Homo sapiens; RNA-Seq
library_construction_protocol: Firstly, plasma isolation was performed from blood samples before and 1 week after treatment. For this purpose, samples taken in PAXgene® Blood RNA Tube were centrifuged at 4000 rpm for 15 minutes and plasma was stored at -80 o . Then, exo-RNA isolation was performed using QIAGEN exoRNeasy Midi kit (Qiagen, Hilden, Germany). For exo isolation, exo bound with XBP were collected by passing PBS through the column, and characterization experiments were carried out. 1 ml plasma samples were transferred to 15 ml conical tubes and RNA isolation was performed. The plasma sample was mixed with 2X binding buffer (XBP) at a ratio of 1:1 and added to the exoEasy membrane affinity column. Then, the washed samples were lysed by adding QIAzol. The lysate was collected after centrifugation at 500 g and after adding chloroform, the aqueous phase was recovered and mixed with ethanol. The samples were added to the RNeasy MinElute spin column, centrifuged and, eluted with 30 µl of RNase- free water after washing To perform RNA sequencing, mRNA and small RNA libraries were generated using the NGS sequencing Illumina NextSeq 500 platform with the SMARTer smRNA-Seq Kit (Illumina, Takara Bio, Shiga, Japan) according to the manufacturer's instructions. The libraries were gel-purified and assessed using the D1000 ScreenTape System (Agilent Technologies, Waldbronn, Germany).
identifiers.namespace: GEO
identifiers.id: GSM8806078_r1