Illumina HiSeq 2000 sequencing; Transcriptomics of Arabidopsis thaliana wild-type and epimutants under control or long-term heat stress
Transcriptomics of Arabidopsis thaliana wild-type and epimutants under control or long-term heat stress
At least 50 seedlings were pooled from three independent petri dishes for RNA extraction. Arabidopsis thaliana seeds were surfaced sterilized and sown in petri dishes with agarified (0.8% Bacto Agar) 0.5x MS medium, containing 1% sucrose and 0.05% MES at pH 5.7. After stratification for 3 days at 4°C, seedlings were grown in a CU-22L growth chamber (Percival) under 21°C and a 12/12 h (day/night) light cycle.  For treatment, 7 days-old seedlings were subjected to changes in temperature only during the light period. Long-term heat consisted of an entrainment treatment of 5 heat cycles (day 37C, night 21C, 12/12 h; age at harvest 12 days). Controls were kept at 21C (age at harvest 12 days). Total RNA was isolated from biologically duplicated samples using the the Plant-RNAeasy kit (Invitrogen), following provider's instructions. Libraries were prepared with the corresponding Illumina library preparation kit, following provider's instructions.