Assessment of direct versus indirect magnetic bead-based T-cell isolation procedures followed by magnetic bead-based DNA isolation

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. Pur...

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Autores principales: Rosenbaum, Anna, Bleck, Ellen, Schneider, Matthias, Pongratz, Georg, Vordenbäumen, Stefan
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BMJ Publishing Group 2016
Materias:
Letter to the Editor
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4964240/
https://www.ncbi.nlm.nih.gov/pubmed/27547441
http://dx.doi.org/10.1136/lupus-2016-000167
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author Rosenbaum, Anna
Bleck, Ellen
Schneider, Matthias
Pongratz, Georg
Vordenbäumen, Stefan
author_facet Rosenbaum, Anna
Bleck, Ellen
Schneider, Matthias
Pongratz, Georg
Vordenbäumen, Stefan
author_sort Rosenbaum, Anna
collection PubMed
description 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.
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spelling pubmed-49642402016-08-19 Assessment of direct versus indirect magnetic bead-based T-cell isolation procedures followed by magnetic bead-based DNA isolation Rosenbaum, Anna Bleck, Ellen Schneider, Matthias Pongratz, Georg Vordenbäumen, Stefan Lupus Sci Med Letter to the Editor 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. BMJ Publishing Group 2016-07-19 /pmc/articles/PMC4964240/ /pubmed/27547441 http://dx.doi.org/10.1136/lupus-2016-000167 Text en Published by the BMJ Publishing Group Limited. For permission to use (where not already granted under a licence) please go to http://www.bmj.com/company/products-services/rights-and-licensing/ 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/
spellingShingle Letter to the Editor
Rosenbaum, Anna
Bleck, Ellen
Schneider, Matthias
Pongratz, Georg
Vordenbäumen, Stefan
Assessment of direct versus indirect magnetic bead-based T-cell isolation procedures followed by magnetic bead-based DNA isolation
title Assessment of direct versus indirect magnetic bead-based T-cell isolation procedures followed by magnetic bead-based DNA isolation
title_full Assessment of direct versus indirect magnetic bead-based T-cell isolation procedures followed by magnetic bead-based DNA isolation
title_fullStr Assessment of direct versus indirect magnetic bead-based T-cell isolation procedures followed by magnetic bead-based DNA isolation
title_full_unstemmed Assessment of direct versus indirect magnetic bead-based T-cell isolation procedures followed by magnetic bead-based DNA isolation
title_short Assessment of direct versus indirect magnetic bead-based T-cell isolation procedures followed by magnetic bead-based DNA isolation
title_sort assessment of direct versus indirect magnetic bead-based t-cell isolation procedures followed by magnetic bead-based dna isolation
topic Letter to the Editor
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4964240/
https://www.ncbi.nlm.nih.gov/pubmed/27547441
http://dx.doi.org/10.1136/lupus-2016-000167
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