Figure 7

High throughput validation of novel alternative splicing features. From the novel features identified in the analyses of the human retinal transcriptome, 14,696 novel exons, exon skipping, and alternate 3′/5′ splice events were selected for capture using a custom Agilent SureSelect RNA Capture System. RNA samples from human retina, brain, muscle, and liver were enriched using baits designed against the novel features. The enriched samples were sequenced and aligned to the genome using the RUM pipeline. Following alignment, the samples were analyzed for the detection of the novel and annotated transcripts associated with the putative novel feature. A) 99% of the novel features detected by the RNA-Seq analyses were detected by the RNA capture in the individual retinal samples. 93% of all novel features were detected in a pooled library prepared from all three retinal RNA samples. Fewer of the novel transcript features were detected in RNA brain (71%), muscle (58%), and liver (61%). B) 6696 novel features were detected in all 6 RNA samples, while 1968 were detected only in the retina samples.