GSM4191802: ZAT14-like_14_hours_1; Arabidopsis thaliana; RNA-Seq
Col-0 seedlings carrying pWOL[xve]::ZAT14-mTurq and pWOL[xve]::ZAT14like-mTurq constructs were transferred to ½ MS plates supplemented with B-estradiol (5uM final concentration) 10 and 14 hours prior sampling the root meristems for RNA extraction. After induction, 5 days old seedlings were transferred to 1/2 MS medium plates with 5% agar in batches of 15 for easier dissection. Root meristems were cut off with a syringe needle and immediately transferred to RNAlater solution. All meristems were collected in total volume of 20ul of RNAlater. Total RNAs were extracted using RNeasy Mini Kit (Qiagen) according to manufacturer's instructions.  Library preparation and sequencing was done by Novogene (beijing, China) as follows: After the QC procedures, mRNA from eukaryotic organisms is enriched using oligo(dT) beads. For prokaryotic samples, rRNA is removed using a specialized kit that leaves the  mRNA. The mRNA from either eukaryotic or prokaryotic sources is then fragmented randomly in fragmentation buffer, followed by cDNA synthesis using random hexamers and  reverse transcriptase. After first-strand synthesis, a custom second-strand synthesis buffer (Illumina) is added with dNTPs, RNase H and Escherichia coli polymerase I to generate the  second strand by nick-translation. The final cDNA library is ready after a round of purification, terminal repair, A-tailing, ligation of sequencing adapters, size selection and PCR  enrichment.
GEO Accession
GSM4191802