organism: Arabidopsis thaliana
description: Protocols: The Arabidopsis seedlings grown on MS media for 2 weeks was treated with 2μM t-zeatin for 30 minutes and then total RNA was extracted from Arabidopsis seedlings using TRIzol reagent (Invitrogen) . The total RNA samples are first treated with DNase I to degrade any possible DNA contamination. Then the mRNA is enriched by using the oligo(dT) magnetic beads (for eukaryotes) or by removing rRNAs from the total RNA (for prokaryotes). Mixed with the fragmentation buffer, the mRNA is fragmented into short fragments (about 200 bp). Then the first strand of cDNA is synthesized by using random hexamer-primer. Buffer, dNTPs, RNase H and DNA polymerase I are added to synthesize the second strand. The double strand cDNA is purified with magnetic beads. End reparation and 3'-end single nucleotide A (adenine) addition is then performed. Finally, sequencing adaptors are ligated to the fragments. The fragments are enriched by PCR amplification. During the QC step, Agilent 2100 Bioanaylzer and ABI StepOnePlus Real-Time PCR System are used to qualify and quantify of the sample library. The library products are ready for sequencing via Illumina HiSeqTM 2000 or other sequencer when necessary.
attributes.tag: ENA first public
attributes.value: 2016-06-17
attributes.tag: ENA last update
attributes.value: 2016-06-07
attributes.tag: External Id
attributes.value: SAMEA4028679
attributes.tag: INSDC center alias
attributes.value: Institute of Botany, Chinese Academy of Sciences
attributes.tag: INSDC center name
attributes.value: Institute of Botany, Chinese Academy of Sciences
attributes.tag: INSDC first public
attributes.value: 2016-06-17T17:01:49Z
attributes.tag: INSDC last update
attributes.value: 2016-06-07T21:57:21Z
attributes.tag: INSDC status
attributes.value: public
attributes.tag: Submitter Id
attributes.value: E-MTAB-4603:deg9-ctk
attributes.tag: broker name
attributes.value: ArrayExpress
attributes.tag: common name
attributes.value: thale cress
attributes.tag: genotype
attributes.value: deg9 mutant
attributes.tag: sample name
attributes.value: E-MTAB-4603:deg9-ctk