Bibliographic Details
| Title: |
Preparation of optimized concanavalin A-conjugated Dynabeads® magnetic beads for CUT&Tag. |
| Authors: |
Fujiwara, Yasuhiro, Tanno, Yuji, Sugishita, Hiroki, Kishi, Yusuke, Makino, Yoshinori, Okada, Yuki |
| Source: |
PLoS ONE; 11/16/2021, Vol. 16 Issue 11, p1-12, 12p |
| Subject Terms: |
CHIMERIC proteins, MAGNETIC particles, TRANSCRIPTION factors, CHROMATIN |
| Abstract: |
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. [ABSTRACT FROM AUTHOR] |
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| Database: |
Complementary Index |
