title: GSM2354324: P040_3b; Zea mays; RNA-Seq
library_construction_protocol: At the end of the previously described growing period, leaf samples of the inbred lines were collected at the three heat levels with n=10 replications at each of the three heat levels. A sample of about 0.5 cm2 was cut from the centre of the latest fully developed leaf of each plant, immediately frozen in liquid nitrogen, and stored at −80°C. The leaf tissue of five replications was pooled to a total of two replications for each genotype - heat level combination to reduce biological variation for the following RNA sequencing. This resulted in a total of 47 samples (the sample for one replication of a genotype - heat level combination was missing). Total RNA was isolated using the RNeasy Plant Kit (Qiagen, Hilden, Germany). RNA quantity was assessed and quality control was performed using a Qubit fluorometer (Life Technologies, Darmstadt, Germany) and the 2100 Bioanalyzer (Agilent Technologies, Böblingen, Germany). DNA was removed using the TURBO DNA free Kit (Ambion, Kaufungen, Germany) and the solution was purified using the RNeasy Ⓡ MinElute Ⓡ Cleanup spin columns (Qiagen). rRNA was depleted prior to sequencing using the RiboMinus™Plant Kit (Invitrogen, Life Technologies, Darmstadt, Germany). Library preparation and RNA sequencing were performed at the Max Planck Genome Centre Cologne using an Illumina HiSeq2000 sequencing machine (Illumina, Inc., San Diego, CA USA)