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Assessment of direct versus indirect magnetic bead-based T-cell isolation procedures followed by magnetic bead-based DNA isolation
  1. Anna Rosenbaum,
  2. Ellen Bleck,
  3. Matthias Schneider,
  4. Georg Pongratz and
  5. Stefan Vordenbäumen
  1. Medical Faculty, Hiller Research Center Rheumatology at University Hospital Düsseldorf, Heinrich-Heine-University, Düsseldorf, Germany
  1. Correspondence to Dr Stefan Vordenbäumen; Stefan.Vordenbaeumen{at}med.uni-duesseldorf.de

Abstract

Objective To compare direct and indirect bead-based T-cell isolation followed by magnetic bead-based DNA isolation.

Methods T-cells were isolated by direct or indirect selection with magnetic bead coated antbiodies followed by magnetic bead-based automated DNA isolation in 10 healthy subjects. Purity of T-cells, purity of DNA (by A260/A280 ratio measurement) and DNA concentration were assessed.

Results Direct and indirect labelling resulted in comparable T-cell purity (93.11±1.47% vs. 94.99±1.54%, p= 0.125) and DNA concentration per cell (50.97±14.15 ng/(mlxcell) vs. 49.53±13.62 ng/(mlxcell), p=0.492), while DNA purity was significantly higher after direct labelling (1.82±0.05 vs. 1.78±0.03, p=0.0488).

Conclusions Both direct and indirect magnetic bead-based T-cell selection may be used prior to magnetic bead-based DNA isolation procedures.

  • T Cells
  • magnetic beads
  • DNA isolation
  • epigenetics

This is an Open Access article distributed in accordance with the Creative Commons Attribution Non Commercial (CC BY-NC 4.0) license, which permits others to distribute, remix, adapt, build upon this work non-commercially, and license their derivative works on different terms, provided the original work is properly cited and the use is non-commercial. See: http://creativecommons.org/licenses/by-nc/4.0/

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