Quick rna miniprep isolation plus kit

RNA was isolated from PBMC lysates using the Quick RNA MiniPrep Isolation Plus Kit (ZymoResearch, Freiburg, Germany). A miRNeasy serum spike-in miR-39 miRNA (Qiagen, Hilden, Germany) was integrated in the micro-RNA isolation procedure. Equal amounts of RNA (10ng) were reverse transcribed using the Advanced miRNA cDNA Synthesis Kit (Thermo Fisher Scientific). cDNA samples were diluted 1:60, followed by qPCR using the miRCURY LNA SYBR Green PCR Kit (Qiagen). Primers used for the detection of Hsa-142-3p/5p (YP00204291/YP00204722) were purchased from Qiagen. The samples were processed in duplicates on a StepOnePlus Cycler (Thermo Fisher Scientific). Data were normalized by miR-39 and related to healthy control donor miR. Thus, results were expressed as the x-fold difference compared to the healthy control.

RNA was isolated from PBMC lysates using the Quick RNA MiniPrep Isolation Plus Kit (ZymoResearch, Freiburg, Germany). The RNA concentration and quality (260/280 ratio: 1.8 ± 0.1) were tested using the Nanodrop technique (PEQLAB Biotechnologie GmbH, Erlangen, Germany). Equal amounts of RNA (50 ng) were reverse transcribed using the High-Capacity cDNA Reverse Transcription Kit (Thermo Fisher Scientific).
IL-10 (Hs00961622_m1), Cyp1A (Hs01052496_g1), Cyp1B (Hs00164383_m1), IL-6 (Hs00985639_m1), SOCS1 (HS00705164_s1), SOCS3 (HS02330326_s1), and RPL37A (Hs01102345_m1) mRNA expressions were analyzed using TaqMan probes (Thermo Fisher Scientific) using qPCRBIO Probe Mix High-ROX (Nippon Genetics, Düren, Germany). The samples were processed in duplicates on a StepOnePlus Cycler (Thermo Fisher Scientific). Data were normalized by RPL37A and related to healthy control donor RNA. Thus, results were expressed as the x-fold difference compared to the healthy control.