Rna 6000 nano kit for agilent 2100 bioanalyzer

RNA from 2.5 ml whole blood collected in PAXgene tubes was isolated using the PAXgene Blood miRNA Kit (Qiagen, Hilden, Germany) according to the manufacturer's recommendations. RNA concentration and integrity were measured using Nanodrop 2000 spectrophotometer (Thermo Fisher Scientific, Waltham, MA, USA) and RNA 6000 Nano Kit for Agilent 2100 Bioanalyzer (Agilent Technologies, Santa Clara, CA, USA), respectively. RNA was aliquoted and used for the four experimental approaches CoolMPS, BGISEQ, Illumina and reverse transcriptase-quantitative polymerase chain reaction (RT-qPCR) as described below. The study was approved by the ethical committee of the Medical Faculty of the University of Tuebingen (Nr. 90/2009BO2). A list of samples included in the study is available as Supplementary Table S1.

Total RNA was extracted from twelve root and shoot samples using Plant/Fungi Total RNA Purification Kit, according to the manufacturer’s instructions (Cat. 25800, Norgen, Canada) and treated with RNase-Free DNase I (Cat. 25710, Norgen) during RNA purification. RNA purity and quality were assessed using NanoDrop 1000 spectrophotometer whereas its integrity was examined by RNA 6000 Nano Kit for Agilent 2100 Bioanalyzer (Agilent Technologies, Waldbronn, Germany). High quality RNA samples with RIN ≥ 7.2 were sent to Admera Health LLC (New Jersey, USA) for cDNA library construction and sequencing.