GSM3104220: 0_DAT_Heat_1 [small RNA]; Triticum aestivum; ncRNA-Seq
Leaf tissue was collected from all heat-stressed and control individual plants immediately at the end of the stress period, i.e., time point zero day after treatment (0 DAT), and at one and four days after treatment (1 and 4 DAT). Total RNA was isolated from all 432 leaf tissue samples using TRI-reagent (Ambion, Naugatuck, CT) as previously described (Ragupathy et al., 2016; Scientific Reports, 6, 39373). sRNA libraries were constructed from low molecular weight (LMW) RNA isolated from 72 µg of total RNA as described by Lu and Souret (2010) in Methods and protocols (eds. B.C. Meyers & P.J. Green), pp. 107-125. Humana Press, New York. libraries were constructed using Illumina TruSeq® small RNA kit following the manufacturer's instructions. Six pools, each containing equimolar amounts of 12 random libraries and where each library was present in three different pools. The library pools were normalized to 2 nM and sequenced on a MiSeq for 50 cycles using a MiSeq Reagent Kit v2 (Illumina). A total of six runs were performed to sequence the six pools where each library was sequenced in three separate runs.