title: Illumina Genome Analyzer IIx paired end sequencing; RNASeq profiling of AZD8186-treated HCC70 xenografts
design: RNASeq profiling of AZD8186-treated HCC70 xenografts
library_construction_protocol: All animal experiments were performed to the according to the Home Office UK.  HCC70 cells (1x106  cells in RPMI serum free medium mixed 50:50 with MatrigelTM) tumours were implanted in the flank of female nude mice (nu/nu:Alpk) (AstraZeneca, Alderley Park, UK) between the ages of 8 and 12 weeks. Once tumours reached ~200-500mm3 animals were randomised into control and treatment groups. Tumour volume was calculated twice weekly from bilateral caliper measurements using the formula (Length x width x width) x π/6). Dosage of treated xenografts For xenograft models, cell pellets and tissues from xenograft models were snap frozen. Total RNA was extracted using miRNeasy kit (Qiagen), with DNase treatment, following manufacturer's instructions. RNA quantity was assessed by Nanodrop 2000. Samples profiled for RNA-seq were assessed for RNA integrity (RIN>7) using the RNA 6000 Nano Assay on the BioAnalyser (Agilent).  Sample were sent for RNA-sequencing at Liverpool Centre for Genomic Research. Total RNA was ribo-depleted, paired-end ScriptSeq library was performed and sequenced for 100bp/100M reads per sample.
attributes.tag: Experimental Factor: AZD8186
attributes.value: compound
attributes.tag: Experimental Factor: 100
attributes.value: dose