Comparison of RO-seq Isolation Protocols and Library Preparation Methods

The study of Nascent RNA has expanded with the development of run-on sequencing protocols (RO-seq). These protocols are capable of capturing nascent RNAs with remarkable precision, enabling the study of polymerase behavior at high resolution. Many RO-seq protocols have been developed within the past decade; however, the degree to which these protocols capture the same information has not been explored. We provide here a comparison of these different protocols, which reveal significant biases in each protocol.