title: GSM1288410: B. juncea high temperature stressed (BJH); Brassica juncea; RNA-Seq
library_construction_protocol: Tissues obtained from different time points of each stress were pooled and enriched low molecular weight (LMW) RNA was isolated using GITC-LiCl method (Chomczynski and Sacchi, 1987) with some modifications. Tissue was homogenized in liquid nitrogen and total RNA was extracted using GITC buffer and phenol-chloroform purification steps followed by ethanol precipitation. Total RNA was re-precipitated with lithium chloride to extract low molecular weight RNA. Small RNA libraries were prepared utilizing Illumina small RNA sample preparation kit v1 according to manufacturer’s instructions. Briefly, equal amount of enriched-LMW RNA (50 μg) was size fractionated on 15% TBE-urea gel and small RNA fraction (18-40 nt) was excised and purified. RNA adapters were ligated on both ends using T4 RNA ligase followed by cDNA preparation utilizing Superscript II (Invitrogen, USA). The prepared cDNA was PCR amplified and the amplicon of approximately 100 bp was excised from a 6% TBE gel. The quality and quantity of prepared libraries was evaluated utilizing DNA 1000 kit on a bioanalyzer (Agilent, USA).
attributes.tag: GEO Accession
attributes.value: GSM1288410