Preparation of optimized concanavalin A-conjugated Dynabeads® magnetic beads for CUT&Tag.

Epigenome research has employed various methods to identify the genomic location of proteins of interest, such as transcription factors and histone modifications. A recently established method called CUT&Tag uses a Protein-A Tn5 transposase fusion protein, which cuts the genome and inserts adapt...

Descripción completa

Guardado en:

Detalles Bibliográficos
Autores principales:	Yasuhiro Fujiwara, Yuji Tanno, Hiroki Sugishita, Yusuke Kishi, Yoshinori Makino, Yuki Okada
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/a283e84c16654645a3d82d15a78346a4
Etiquetas:	Agregar Etiqueta Sin Etiquetas, Sea el primero en etiquetar este registro!

id	oai:doaj.org-article:a283e84c16654645a3d82d15a78346a4
record_format	dspace
spelling	oai:doaj.org-article:a283e84c16654645a3d82d15a78346a42021-12-02T20:13:02ZPreparation of optimized concanavalin A-conjugated Dynabeads® magnetic beads for CUT&Tag.1932-620310.1371/journal.pone.0259846https://doaj.org/article/a283e84c16654645a3d82d15a78346a42021-01-01T00:00:00Zhttps://doi.org/10.1371/journal.pone.0259846https://doaj.org/toc/1932-6203Epigenome research has employed various methods to identify the genomic location of proteins of interest, such as transcription factors and histone modifications. A recently established method called CUT&Tag uses a Protein-A Tn5 transposase fusion protein, which cuts the genome and inserts adapter sequences nearby the target protein. Throughout most of the CUT&Tag procedure, cells are held on concanavalin A (con A)-conjugated magnetic beads. Proper holding of cells would be decisive for the accessibility of Tn5 to the chromatin, and efficacy of the procedure of washing cells. However, BioMag®Plus ConA magnetic beads, used in the original CUT&Tag protocol, often exhibit poor suspendability and severe aggregation. Here, we compared the BioMag beads and Dynabeads® magnetic particles of which conjugation of con A was done by our hands, and examined the performance of these magnetic beads in CUT&Tag. Among tested, one of the Dynabeads, MyOne-T1, kept excessive suspendability in a buffer even after overnight incubation. Furthermore, the MyOne-T1 beads notably improved the sensitivity in CUT&Tag assay for H3K4me3. In conclusion, the arrangement and the selection of MyOne-T1 refine the suspendability of beads, which improves the association of chromatin with Tn5, which enhances the sensitivity in CUT&Tag assay.Yasuhiro FujiwaraYuji TannoHiroki SugishitaYusuke KishiYoshinori MakinoYuki OkadaPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 16, Iss 11, p e0259846 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Yasuhiro Fujiwara Yuji Tanno Hiroki Sugishita Yusuke Kishi Yoshinori Makino Yuki Okada Preparation of optimized concanavalin A-conjugated Dynabeads® magnetic beads for CUT&Tag.
description	Epigenome research has employed various methods to identify the genomic location of proteins of interest, such as transcription factors and histone modifications. A recently established method called CUT&Tag uses a Protein-A Tn5 transposase fusion protein, which cuts the genome and inserts adapter sequences nearby the target protein. Throughout most of the CUT&Tag procedure, cells are held on concanavalin A (con A)-conjugated magnetic beads. Proper holding of cells would be decisive for the accessibility of Tn5 to the chromatin, and efficacy of the procedure of washing cells. However, BioMag®Plus ConA magnetic beads, used in the original CUT&Tag protocol, often exhibit poor suspendability and severe aggregation. Here, we compared the BioMag beads and Dynabeads® magnetic particles of which conjugation of con A was done by our hands, and examined the performance of these magnetic beads in CUT&Tag. Among tested, one of the Dynabeads, MyOne-T1, kept excessive suspendability in a buffer even after overnight incubation. Furthermore, the MyOne-T1 beads notably improved the sensitivity in CUT&Tag assay for H3K4me3. In conclusion, the arrangement and the selection of MyOne-T1 refine the suspendability of beads, which improves the association of chromatin with Tn5, which enhances the sensitivity in CUT&Tag assay.
format	article
author	Yasuhiro Fujiwara Yuji Tanno Hiroki Sugishita Yusuke Kishi Yoshinori Makino Yuki Okada
author_facet	Yasuhiro Fujiwara Yuji Tanno Hiroki Sugishita Yusuke Kishi Yoshinori Makino Yuki Okada
author_sort	Yasuhiro Fujiwara
title	Preparation of optimized concanavalin A-conjugated Dynabeads® magnetic beads for CUT&Tag.
title_short	Preparation of optimized concanavalin A-conjugated Dynabeads® magnetic beads for CUT&Tag.
title_full	Preparation of optimized concanavalin A-conjugated Dynabeads® magnetic beads for CUT&Tag.
title_fullStr	Preparation of optimized concanavalin A-conjugated Dynabeads® magnetic beads for CUT&Tag.
title_full_unstemmed	Preparation of optimized concanavalin A-conjugated Dynabeads® magnetic beads for CUT&Tag.
title_sort	preparation of optimized concanavalin a-conjugated dynabeads® magnetic beads for cut&tag.
publisher	Public Library of Science (PLoS)
publishDate	2021
url	https://doaj.org/article/a283e84c16654645a3d82d15a78346a4
work_keys_str_mv	AT yasuhirofujiwara preparationofoptimizedconcanavalinaconjugateddynabeadsmagneticbeadsforcuttag AT yujitanno preparationofoptimizedconcanavalinaconjugateddynabeadsmagneticbeadsforcuttag AT hirokisugishita preparationofoptimizedconcanavalinaconjugateddynabeadsmagneticbeadsforcuttag AT yusukekishi preparationofoptimizedconcanavalinaconjugateddynabeadsmagneticbeadsforcuttag AT yoshinorimakino preparationofoptimizedconcanavalinaconjugateddynabeadsmagneticbeadsforcuttag AT yukiokada preparationofoptimizedconcanavalinaconjugateddynabeadsmagneticbeadsforcuttag
_version_	1718374841581568000

Preparation of optimized concanavalin A-conjugated Dynabeads® magnetic beads for CUT&Tag.

Ejemplares similares