title: Illumina HiSeq 2000 paired end sequencing; Yeast transcriptome profiling in replicative ageing
design: Yeast transcriptome profiling in replicative ageing
library_construction_protocol: The cells were grown in yeast nitrogen base (YNB) without amino acids (ForMedium, Norfolk, UK) supplemented with 2% glucose, at a temperature of 30˚C, unless indicated differently. Precultures in flasks were shaken at 300 RPM. ). The quality and integrity of the RNA-samples was determined with a Nanodrop ND1000 spectrophotometer and analyzed on a RNA 6000 Lab-on-a-Chip using bioanalyzer (Agilent Technologies, Santa Clara, CA, USA). The cDNA libraries were generated by using the Illumina TruSeq mRNA-Seq Sample Prep Kit v2 (Illumina, San Diego, CA, USA). In short, mRNA was isolated from total RNA using the oligo-dT-magnetic beads, fragmented and cDNA synthesis was performed. The cDNA was ligated with the sequencing adapters and amplified by PCR. The quality of the amplified cDNA was measured with a DNA 1000 Lab-on-a-Chip. The fragment sizes ranged between 300 and 500 bp.
attributes.tag: Experimental Factor: age
attributes.value: 0 hour
attributes.tag: Experimental Factor: mix
attributes.value: 2
attributes.tag: LIBRARY_STRAND
attributes.value: not applicable