Partial bisulfite conversion for unique template sequencing.
Nucleic Acids Res
; 46(2): e10, 2018 01 25.
Article
en En
| MEDLINE
| ID: mdl-29161423
ABSTRACT
We introduce a new protocol, mutational sequencing or muSeq, which uses sodium bisulfite to randomly deaminate 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 135 000 restriction fragments in a 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.
Texto completo:
1
Colección:
01-internacional
Banco de datos:
MEDLINE
Asunto principal:
Sulfitos
/
Moldes Genéticos
/
ADN
/
Secuenciación de Nucleótidos de Alto Rendimiento
Idioma:
En
Revista:
Nucleic Acids Res
Año:
2018
Tipo del documento:
Article