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...

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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/3254e3df616847a6b2461867e6b27031
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spelling	oai:doaj.org-article:3254e3df616847a6b2461867e6b270312021-11-25T06:10:51ZPreparation of optimized concanavalin A-conjugated Dynabeads® magnetic beads for CUT&Tag1932-6203https://doaj.org/article/3254e3df616847a6b2461867e6b270312021-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC8594809/?tool=EBIhttps://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 (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/3254e3df616847a6b2461867e6b27031
work_keys_str_mv	AT yasuhirofujiwara preparationofoptimizedconcanavalinaconjugateddynabeadsmagneticbeadsforcuttag AT yujitanno preparationofoptimizedconcanavalinaconjugateddynabeadsmagneticbeadsforcuttag AT hirokisugishita preparationofoptimizedconcanavalinaconjugateddynabeadsmagneticbeadsforcuttag AT yusukekishi preparationofoptimizedconcanavalinaconjugateddynabeadsmagneticbeadsforcuttag AT yoshinorimakino preparationofoptimizedconcanavalinaconjugateddynabeadsmagneticbeadsforcuttag AT yukiokada preparationofoptimizedconcanavalinaconjugateddynabeadsmagneticbeadsforcuttag
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Preparation of optimized concanavalin A-conjugated Dynabeads® magnetic beads for CUT&Tag

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