Single-Molecule Analysis of Pre-mRNA Splicing with Colocalization Single-Molecule Spectroscopy (CoSMoS).
Methods Mol Biol
; 1648: 27-37, 2017.
Article
en En
| MEDLINE
| ID: mdl-28766287
ABSTRACT
Recent development of single-molecule techniques to study pre-mRNA splicing has provided insights into the dynamic nature of the spliceosome. Colocalization single-molecule spectroscopy (CoSMoS) allows following spliceosome assembly in real time at single-molecule resolution in the full complexity of cellular extracts. A detailed protocol of CoSMoS has been published previously (Anderson and Hoskins, Methods Mol Biol 1126217-241, 2014). Here, we provide an update on the technical advances since the first CoSMoS studies including slide surface treatment, data processing, and representation. We describe various labeling strategies to generate RNA reporters with multiple dyes (or other moieties) at specific locations.
Palabras clave
Texto completo:
1
Banco de datos:
MEDLINE
Asunto principal:
Precursores del ARN
/
Empalme del ARN
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Imagen Molecular
Tipo de estudio:
Guideline
Límite:
Animals
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Humans
Idioma:
En
Año:
2017
Tipo del documento:
Article