GSM2480510: Control_bio_rep2_tech_rep4; Lobosphaera incisa; RNA-Seq
Total RNA was isolated from samples collected at the specified time-points: time 0, 12h under HL, 72h under NL and 72h under HL. Lobosphaera incisa biomass of a total amount of approx. 15-20 mg (on the basis of DW measurements) was harvested by centrifugation in 2 ml safe-lock eppendorf tubes. The cell pellet was immediately frozen in liquid nitrogen. Cells were broken in the mixer mill MM 400 (Retsch, Germany) with the addition of 2 and 5 mm iron beads using 7-8 cycles of 45 sec with intermediate freezing in liquid nitrogen between the cycles. Broken cell powder was immediately used for RNA isolation with the SV Total RNA isolation System (Promega Corporation, USA) according to the user’s manual instructions. Total RNA concentration was measured by a NanoDrop 1000 Spectrophotometer (Thermo Fisher Scientific Inc., USA) and calculated in the NanoDrop software, 3.0.0 (Coleman Technologies Inc., USA). The RNA integrity was estimated on the formaldehyde gel electrophoresis and on a 2100 Electrophoresis Bioanalyzer (Agilent technologies). RNA libraries were prepared for sequencing using Illumina Truseq protocol (50-nt single-end reads and 100-nt paired-end reads, insert size ~150 bp). Sequencing was done on a on a HiSeq 1000 instrument. Library preparation and sequencing was performed by the transcriptomics platform of the Institut de Biologie de l'École Normale Supérieure, Paris, France.
GEO
GSM2480510
GEO Accession
GSM2480510