title: Sample 5
organism: Arabidopsis thaliana
description: Protocols: Two-week-old seedlings were harvested and flash frozen in liquid nitrogen. Two-week-old seedlings grown in 2 ml liquid 0.5x MS salts with 0.5% (w/v) sucrose in six-well plate (Greiner Bio-One) in a growth chamber at 12-h light(120 µmol photons m-2 s-1, 22°C) and 12-h dark (18°C; CLF Plant Climatics, Emersacker, Germany). Total RNA was isolated by RNeasy Plant Mini Kit (Qiagen). mRNA was enriched using oligo(dT) bead. The mRNA is fragmented randomly by adding fragmentation buffer, then the cDNA is synthesized by using mRNA template and random hexamers primer, after which a custom second-strand synthesis buffer (Illumina) , dNTPs, RNase H and DNA polymerase I are added to initiate the second-strand synthesis. Second, after a series of terminal repair, A ligation and sequencing adaptor ligation, the double-stranded cDNA library is completed through size selection and PCR enrichment.
attributes.tag: isolate
attributes.value: not applicable
attributes.tag: cultivar
attributes.value: Col-0
attributes.tag: organism
attributes.value: Arabidopsis thaliana
attributes.tag: organism
attributes.value: Arabidopsis thaliana
attributes.tag: collection date
attributes.value: not collected
attributes.tag: scientific_name
attributes.value: Arabidopsis thaliana
attributes.tag: common name
attributes.value: thale cress
attributes.tag: organism part
attributes.value: whole plant
attributes.tag: age
attributes.value: 2
attributes.tag: genotype
attributes.value: dph2-1
attributes.tag: geographic location (country and/or sea)
attributes.value: not collected