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Protocol to measure centromeric array size changes using droplet digital PCR-based quantification of higher-order repeats.
Showman, Soyeon; Talbert, Paul B; Xu, Yiling; Henikoff, Steven.
Afiliación
  • Showman S; Basic Sciences Division, Fred Hutchinson Cancer Center, Seattle, WA 98109, USA; Molecular and Cellular Biology Graduate Program, University of Washington, Seattle, WA 98195, USA; Howard Hughes Medical Institute, Chevy Chase, MD 20815, USA.
  • Talbert PB; Basic Sciences Division, Fred Hutchinson Cancer Center, Seattle, WA 98109, USA; Howard Hughes Medical Institute, Chevy Chase, MD 20815, USA.
  • Xu Y; Basic Sciences Division, Fred Hutchinson Cancer Center, Seattle, WA 98109, USA; Howard Hughes Medical Institute, Chevy Chase, MD 20815, USA.
  • Henikoff S; Basic Sciences Division, Fred Hutchinson Cancer Center, Seattle, WA 98109, USA; Howard Hughes Medical Institute, Chevy Chase, MD 20815, USA. Electronic address: steveh@fredhutch.org.
STAR Protoc ; 5(3): 103218, 2024 Sep 20.
Article en En | MEDLINE | ID: mdl-39068651
ABSTRACT
Centromere length changes occurring during somatic cell divisions can be estimated by quantifying the copy numbers (CNs) of higher-order repeats (HORs), which are nested repeats of monomers that comprise centromeric arrays. Here, we present a protocol for single-cell isolation for clonal evolution followed by droplet digital PCR-based quantification. The assay measures HOR CNs across subclones to determine the frequency and degree of changes in HOR CNs. This protocol tests the underlying molecular mechanisms responsible for rapid centromere sequence evolution. For complete details on the use and execution of this protocol, please refer to Showman et al.1.
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Centrómero / Reacción en Cadena de la Polimerasa Límite: Humans Idioma: En Revista: STAR Protoc Año: 2024 Tipo del documento: Article País de afiliación: Estados Unidos Pais de publicación: Estados Unidos
Texto completo
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Imprimir
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Centrómero / Reacción en Cadena de la Polimerasa Límite: Humans Idioma: En Revista: STAR Protoc Año: 2024 Tipo del documento: Article País de afiliación: Estados Unidos Pais de publicación: Estados Unidos