title: GSM5943589: cs_cage_embryo_rep1; Triticum aestivum; RNA-Seq
library_construction_protocol: The seedlings (above-ground parts) in soil were harvested after 9-day growth. The root in Hoagland solution were harvested after 9-day growth. The spikelets at booting stage (Feeke 10) were harvested. The fresh immature embryos (14days post anthesis) were isolated and either frozen in liquid nitrogen For CAGE-seq，total RNA was treated with RQ1 DNase to remove DNA. The total RNA was treated with T4 polynucleotide kinase subsequently digested with Terminator 5´-Phosphate-Dependent Exonuclease to enrich the capped mRNA. Next, reverse transcription was performed with RT primer harboring a 3'-adaptor sequence and randomized hexamer. Subsequently, the 5'adaptor harboring three additional rG at the 3'terminus was added to the RT reaction to allow template switching and tagging of the 5' adaptor. The cDNAs were treated with Exonuclease I to digest the primers, purified and amplified with PCR primers. The libraries were applied to Illumina Novaseq 6000 system for 150 nt paired-end sequencing.