title: 2_l1
organism: Lolium perenne
description: Protocols: Two perennial ryegrass (Lolium perenne L.) genotypes, 'Falster' originated from Denmark, adapted to cold climates and 'Veyo' originated from Italy, adapted to warmer climates were propagated vegetatively in a glasshouse. Following propagation plants were grown in a glasshouse (photoperiod, 9 h : 15 h, light : dark) at ~20 degrees C for 12 weeks. Plants (at the pre-elongation stage) were then transferred to a growth chamber for 7 d (photoperiod, 9 h : 15 h, light : dark) with 450umol photons m-2 s-1 light intensity at 20 degrees C. Plants were cold acclimated in a controlled growth room for 17 d (photoperiod, 9 h : 15 h, light : dark) with 450umol photons m-2 s-1 light intensity and relative humidity of ~70% at 7 degrees C. Green tissue (top) samples were harvested in three biological replicates at the onset of the daily photoperiod at 0, 9, 13 and 17 d of 7 degrees C cold acclimation for RNA-Seq analysis. Samples were frozen in liquid nitrogen immediately after harvesting and stored at -80 degrees C until analysis. Leaf tissues were grounded separately in liquid nitrogen. Total RNA was extracted using RNeasy(r) Plant Mini Kit (Qiagen) and On-column DNAse I digestion (Qiagen) was performed to avoid the genomic DNA contamination according to the manufacturers' instructions. Total RNA quality was verified using an Agilent 2100 Bioanalyzer (RNA 6000 Nano Assay) and quantified by Quant-iTTM RiboGreen(r) RNA Reagent assay following the manufacturers' protocol. Total RNA samples were prepared for sequencing using the TruSeq RNA Sample Preparation Kit (Illumina).
attributes.tag: ENA first public
attributes.value: 2016-04-20
attributes.tag: ENA last update
attributes.value: 2014-07-31
attributes.tag: External Id
attributes.value: SAMEA2677098
attributes.tag: INSDC center alias
attributes.value: Researcher
attributes.tag: INSDC center name
attributes.value: Researcher
attributes.tag: INSDC first public
attributes.value: 2016-04-20T17:01:17Z
attributes.tag: INSDC last update
attributes.value: 2014-07-31T23:20:40Z
attributes.tag: INSDC status
attributes.value: public
attributes.tag: Submitter Id
attributes.value: E-MTAB-2779:2_l1
attributes.tag: broker name
attributes.value: ArrayExpress
attributes.tag: genotype
attributes.value: Veyo
attributes.tag: organism part
attributes.value: leaf
attributes.tag: replicate
attributes.value: 2
attributes.tag: sample name
attributes.value: E-MTAB-2779:2_l1
attributes.tag: time for cold acclimation
attributes.value: 0 day