TY - JOUR T1 - Mutational sequencing for accurate count and long-range assembly JF - bioRxiv DO - 10.1101/149740 SP - 149740 AU - Vijay Kumar AU - Julie Rosenbaum AU - Zihua Wang AU - Talitha Forcier AU - Michael Ronemus AU - Michael Wigler AU - Dan Levy Y1 - 2017/01/01 UR - http://biorxiv.org/content/early/2017/06/13/149740.abstract N2 - We introduce a new protocol, mutational sequencing or muSeq, which randomly deaminates unmethylated cytosines at a fixed and tunable rate. The muSeq protocol marks each initial template molecule with a unique mutation signature that is present in every copy of the template, and in every fragmented copy of a copy. In the sequenced read data, this signature is observed as a unique pattern of C-to-T or G-to-A nucleotide conversions. Clustering reads with the same conversion pattern enables accurate count and long-range assembly of initial template molecules from short-read sequence data. We explore count and low-error sequencing by profiling a 135,000 fragment PstI representation, demonstrating that muSeq improves copy number inference and significantly reduces sporadic sequencer error. We explore long-range assembly in the context of cDNA, generating contiguous transcript clusters greater than 3,000 bp in length. The muSeq assemblies reveal transcriptional diversity not observable from short-read data alone. ER -