GSM1922913: L. lactis TOMSC161_late stationary_6; Lactococcus lactis subsp. lactis; RNA-Seq
25 mL of cultures were harvested at the early and late stationary phase. One volume of ice-cold K-Buffer (20 mM Tris-HCl pH 7.5; 5 mM MgCl2; 20 mM NaN3) was then immediately added to two volumes of each cell suspension and gently mixed. Pellets were collected by centrifugation (15,000 g, 5 min, 4°C), rapidly frozen in liquid nitrogen and stored at -80°C. For the extraction of total RNA, the previous method described by Sperandio et al. (2005) was used. Ribosomal RNA was then depleted using the Ribo-Zero Magnetic Kit (Epicentre; Madison, WI, USA) according to the manufacturer’s instructions. Finally, RNA concentration was quantified at 260 nm using a NanoDrop ND-1000 spectrophotometer (NanoDrop Technologies; Wilmington, DE, USA) and RNA integrity was assessed by gel electrophoresis using an RNA 6000 NANO chip with the 2100 Bioanalyzer (Agilent; Palo Alto, CA, USA) according to the manufacturer’s instructions. RNA-Seq libraries were constructed on cBot (Illumina; San Diego, CA, USA) using TruSeq SR Cluster Kit v3-cBot-HS (Illumina) and a single-end sequencing was performed on Hiseq 1000 (Illumina) using TruSeq SBS Kit v3-HS (Illumina) according to the manufacturer’s instructions.
GEO Accession
GSM1922913