GSM2414987: WW_d1_ZT17-rep2; Brassica rapa; RNA-Seq
Leaf tissue was homogenized and resuspended in LBB buffer (100mM Tris-Cl pH7.5, 500mM LiCl, 10mM EDTA, 1% LiDs, 5mM DTT, 2-BME). mRNA was extracted directly from tissue lysate using Life Technologies Dynabeads oligo (dT)25 (cat. no. 61002), and DNase treated using ThermoFisher TURBO DNase (cat. no. AM2238) in the presence of EDTA. The concentration of mRNA was assessed using a Qubit (Invitrogen, ThermoFisher) and 55ng was used for the cDNA library preparation. The quality of the mRNA was assessed using the Agilent Technologies 2100 Bioanalyzer (Agilent), all RIN values were below 3. The cDNA libraries were made according to the protocol in Wang et al. 2011 (PLoS One 6(10): e26426, doi:10.1371/journal.pone.0026426). The quality and size of the cDNA libraries were assessed using the Agilent Technologies 2100 Bioanalyzer (Agilent) and run on a gel. All library sizes were ~250bp resulting in insert sizes ~150bp with PE100 sequencing. Libraries were multiplexed 12-samples in a lane - well-watered rep1 timepoints 1-12, well-watered rep2 timepoints 1-12, drought rep1 timepoints 1-12, drought rep2 timepoints 1-12.
GEO Accession
GSM2414987