Diagenode

Enhancing single-cell ATAC sequencing with formaldehyde fixation, cryopreservation, and multiplexing for flexible analysis


Tobias Hohl et al.

The assay for transposase-accessible chromatin using sequencing (ATAC-seq) revolutionized the field of epigenetics since its emergence by providing a means to uncover chromatin dynamics and other factors affecting gene expression. The development of single-cell (sc) applications in recent years led to an even deeper understanding of cell type specific gene regulatory mechanisms. One of the major challenges while running ATAC-seq experiments, bulk or sc, is the need for freshly collected cells for successful experiments. While various freezing methods have already been tested and established for bulk and sc ATAC-seq, quality metrics for preserved cells are rather poor or dependent on sampling time when compared to fresh samples. This makes it difficult to conduct all sorts of complex experiments i.e. with multiple conditions, patients, or time course studies. Especially, accounting for batch effects can be difficult if samples need to be processed at different time points of collection. We tackled this issue by adding a fixation step prior to the freezing method. The additional fixation step improved library quality and yield data comparable to fresh samples. The workflow was also tested on multiplexed sc ATAC experiments, set-up for cost-efficient low input sample handling. Sample cross-in, typically encountered in Tn5-based multiplex approaches, were tackled with a computational procedure specifically developed for this approach.

Tags
Tagmentase

Share this article

Published
November, 2024

Source

Products used in this publication

  • Tubes
    C01070010-10
    Tagmentase
  • Kit icon
    C01019043
    Tagmentation Buffer (2x)
  • default alt
    C01070011
    Tagmentase Dilution Buffer

活动

  • American Association of Cancer Research (AACR)
    Chicago (IL), USA
    Apr 25-Apr 30, 2025
  • 14th International Symposium on Minimal Residual Cancer
    Nice, France
    May 7-May 9, 2025
 查看所有活动

 


       Site map   |   Contact us   |   Conditions of sales   |   Conditions of purchase   |   Privacy policy