title: Illumina HiSeq 4000 paired end sequencing; The diurnal transcriptional landscape of the microalga Tetradesmus obliquus
design: The diurnal transcriptional landscape of the microalga Tetradesmus obliquus
library_name: Sample 72_p
library_construction_protocol: After steady state was reached, 8mL samples were taken for RNA extraction (approximately 10 mgDW). Cells were immediately collected by centrifugation (4255 xg, 0°C for 5 min), supernatant was discarded and pellets were frozen in liquid nitrogen and stored at -80°C until further extraction. Samples for RNA extraction were taken in intervals of 1h for WT or every 3h for slm1. 16:8h diurnal cycle adapted cells RNA extraction was performed using the Maxwell® 16 LEV simplyRNA Tissue kit (Promega). Frozen algae pellet (≈200 μL) were submerged in 400 μL of homogenizing buffer supplemented with 8 μL 1-thioglycerol in a 2 mL Lysing matrix C tube (MP), prefilled with a mix of glass beads. Samples were disrupted using a FastPrep-24 instrument (MP). After disruption, all liquid was transferred to a LEV RNA Cartridge. 300 μL lysis buffer were added and the rest of the extraction was performed using a Maxwell MDx AS3000 machine (Promega) following manufacturer's instructions. RNA integrity and quantity were assessed with an Experion system (Bio-Rad), and only high quality samples (RIN value ≥ 7) were selected. Random library
attributes.tag: Experimental Factor: time
attributes.value: 13
attributes.tag: Experimental Factor: environmental history
attributes.value: sample taken during the light
attributes.tag: Experimental Factor: genotype
attributes.value: slm1 homozygous mutant