Next Generation Sequencing Facilitates Quantitative Analysis of Retinoblastoma Transcriptomes

Methods: mRNA profiles of retinoblastoma samples and para-tumor were generated by deep sequencing, in triplicate, using Illumina. The sequence reads that passed quality filters were analyzed at the transcript isoform level with two methods: Burrows–Wheeler Aligner (BWA) followed by ANOVA (ANOVA) and TopHat followed by Cufflinks. qRT–PCR validation was performed using TaqMan and SYBR Green assays Overall design: Retinoblastoma mRNA of retinoblastoma were generated by deep sequencing, using Illumina.