Thursday, 3 June 2021

Single Cell Analysis - latest papers using DRAQ7

DRAQ™ dyes have become a standard tool in FACS sample preparation for downstream molecular analysis (e.g. snATAC, GWAS, RNAseq, etc.) 

Here are recent references to DRAQ7™:

Single cell suspensions were generated from fresh prostate tissue and prepared for BD Biosciences Rhapsody analysis with Calcein AM and DRAQ7™ used to evaluate dead/damaged cell numbers prior to the cell sort.  This work uncovered a transcriptomic remodelling associated with disease progression. 

Chen, Sujun, et al. "Single-cell analysis reveals transcriptomic remodellings in distinct cell types that contribute to human prostate cancer progression." Nature Cell Biology 23.1 (2021): 87-98.

In pursuit of a fuller understanding of the regulators of differentiation of the neutrophil lineage pathway Schwaber et al. (Univ. of Queensland / Avectas / Bluerock Therapeutics) sorted cells for single cell transcriptomics and differentiated progenitors to granulocytes-macrophage cultures for a timed series of proteomic analyses.

The sorting panel was composed of antibody-conjugated fluorophores PE, PerCP-Cy5.5, FITC/AF488, PacificBlue, and BV421. Dead cells were excluded using DRAQ7™. 

Gated IL-3Rα low,CD45RA+ GMP populations were isolated for RNA extraction (Day 0) and the remaining GMPs were used for initiating granulocyte cultures.

Schwaber, Jessica L., et al. "Network mapping of primary CD34+ cells by Ampliseq based whole transcriptome targeted resequencing identifies unexplored differentiation regulatory relationships." PloS one 16.2 (2021): e0246107.

An unprecedented and major study to map the accessible chromatin of 25 different tissues in the human body has been carried out by a team from UCSD and Ludwig Inst for Cancer Research.  They optimised the harvesting of nuclei from each tissue sample and sorted nuclei to enable snATAC sequencing, gathering data on 470,000+ individual nuclei and categorized these into 54 major cell types.  To trigger the sorting of single nuclei for downstream analysis, they were labeled with DRAQ7™.

Zhang, Kai, et al. "A cell atlas of chromatin accessibility across 25 adult human tissues." bioRxiv (2021). DOI:10.1101/2021.02.17.431699

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Related BioStatus blogposts and white papers:

Myth-busting DNA dyes and single cell sorts (September 2023)

Quick and Gentle Nuclei Prep and Sorting (June 2021)

Sample Prep for Single Cell Transcriptomics & Genomics (November 2019)

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