E-MTAB-2349:12053030
Illumina HiSeq 2000 paired end sequencing; heat shock SUMO-2 RNA-seq
heat shock SUMO-2 RNA-seq
43-4
Human osteosarcoma (U2OS) cells were obtained from CRUK and grown at 37C according to standard cell culture protocols. U2OS cells were grown at 37C and either left untreated or incubated at 43C for 4 hours to induce heat shock. Qiagen RNeasy mini kit according to manufacturer's instructions including DNaseI digestion step Isolated total RNA was incubated with DNaseI to digest residual genomic DNA. Poly-A mRNA was purified from the total RNA sample using oligo(dT) coated magnetic beads and eluted with Tris-HCl and heating. Isolated mRNA was fragmented and used for reverse transcription of cDNA (first and second strand cDNA synthesis). Purified cDNA was subjected to end-repair and A-tailing followed by adaptor ligation. Adaptor-ligated cDNA fragments were then PCR amplified and quality and quantity of the cDNA library was validated on an Agilent 2100 Bioanalyzer and ABI StepOnePlus Real-time PCR system, respectively.
CENTRE FOR GENE REGULATION AND EXPRESSION, UNIVERSITY OF DUNDEE, DUNDEE DD1 5EH, UNITED KINGDOM
E-MTAB-2349:12053030
Centre for Gene Regulation and Expression, University of Dundee, Dundee DD1 5EH, United Kingdom
E-MTAB-2349:12053030
Experimental Factor: growth condition
heat shock