Comparison of manual and automated DNA purification for measuring TREC in dried blood spot (DBS) samples with qPCR

Pierre Olivier Lang, Sheila Govind, Moustapha Dramé, Richard Aspinall

Research output: Contribution to journalArticle

11 Citations (Scopus)

Abstract

Automated nucleic acid extractions from dried blood spot (DBS) samples promises standardized sample treatment, low error rates, avoidance of contamination and requirement of less hands-on time. In the present study, non-automated and automated column based extraction processes using the QIAamp Investigator procedure were compared for the extraction of DNA from DBS samples. The concentration and the purity of DNA generated were determined by optical density readings. Furthermore qPCR downstream applications using the nucleic acids extracted with the two processes and albumin and T-cell receptor excision circles (TREC) copy numbers were measured and compared. The influence of the time of storage was also investigated by analyzing samples freshly dried and stored up to 11. weeks at - 20 °C from the same individual. Finally, we provide arguments of preferentially choosing the automated procedure for extracting DNAs from DBS samples when downstream qPCR applications are required.

Original languageEnglish
Pages (from-to)118-127
Number of pages10
JournalJournal of Immunological Methods
Volume384
Issue number1-2
Early online date31 Jul 2012
DOIs
Publication statusPublished - 31 Oct 2012
Externally publishedYes

Keywords

  • DNA extraction
  • Dried blood spots
  • Quantitative PCR
  • Real time PCR
  • Thymic TREC
  • TREC ratio

ASJC Scopus subject areas

  • Immunology and Allergy
  • Immunology

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