Sample 7
Arabidopsis thaliana
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.
isolate
not applicable
cultivar
Col-0
organism
Arabidopsis thaliana
organism
Arabidopsis thaliana
collection date
not collected
scientific_name
Arabidopsis thaliana
common name
thale cress
organism part
whole plant
age
2
genotype
wild type genotype
geographic location (country and/or sea)
not collected