GSM1197432: Drought-3h_nonpolyA; Arabidopsis thaliana; RNA-Seq
Tissues and seedlings were frozen in liquid nitrogen and immediately used to isolate total RNA (QIAGEN RNeasy Plant Mini Kit), which was then quantified using NanoDrop 1000. The same amount of tissue RNA were mixed. Ribosomal RNAs were deleted from the mixed tissue total RNAs using RiboMinus™ Plant Kit repeated three times. The poly(A)- RNAs of seedling samples were collected by removing poly(A)+ RNAs four times using Oligotex mRNA Mini Kit (QIAGEN). Then rRNAs were removed from poly(A)- RNAs three times as the mixed tissue sample. For the mixed tissue sample, RNA was fragmented and enriched by agarose gel at ~330 nt. Strand-specific cDNA library was constructed by ligating different adapters according to the SMART library construction method (Levin et al. 2010), and then sequenced by Illumina Hisequation 2000 using the 100nt paired-end sequencing protocol. For seedling samples, fragmentation and enrichment of RNAs after poly(A)+ RNAs and rRNAs removal were the same as described for the mixed tissue RNA sequencing. Fragments (200~500nt) were excised from the gel and purified for sequencing. We used the SMART™ cDNA Library Construction Kit for strand-specific cDNA library construction. The 100nt single-end sequencing protocol of Illumina Hisequation 2000 was selected.
GEO Accession
GSM1197432