title: NextSeq 500 paired end sequencing; Transcriptomics of Arabidopsis thaliana wild-type and nrpd1-3 under control or heat stress
design: Transcriptomics of Arabidopsis thaliana wild-type and nrpd1-3 under control or heat stress
library_construction_protocol: A. thaliana seeds were surfaced sterilized and sown in petri dishes with agarified (0.8% Bacto Agar, Becton Dickinson) 0.5x MS medium (Duchefa), containing 1% sucrose and 0.05% MES at pH 5.7. After 3 days of stratification at 4°C, plants were grown in a CU-22L growth chamber (Percival) under 12/12 hours (day/night) light cycle.  For the heat-induction, 7 days old plants grown at 21°C were first placed for 24 hours on top of a chilling platform build-in within the growth chamber and maintained by an external mini-chiller at 4°C, but with the chamber temperature set at 6°C. This was followed by 24 hours with the chamber set at 37°C for the stressed plants, whereas control plants were moved back to 21°C.  RNA was isolated using the Plant-RNAeasy kit (Invitrogen). Strand-specific libraries for RNA-seq were prepared with 2 µg of RNA using the TruSeq Stranded Total RNA Library Prep Kit (Illumina); following the provider's instructions.
attributes.tag: Experimental Factor: wild type
attributes.value: genotype
attributes.tag: Experimental Factor: warm/hot temperature regimen
attributes.value: environmental stress