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

Détails

ID Serval
serval:BIB_7CF41ED25F8E
Type
Article: article d'un périodique ou d'un magazine.
Collection
Publications
Titre
Comparison of manual and automated DNA purification for measuring TREC in dried blood spot (DBS) samples with qPCR.
Périodique
Journal of Immunological Methods
Auteur(s)
Lang P.O., Govind S., Dramé M., Aspinall R.
ISSN
1872-7905 (Electronic)
ISSN-L
0022-1759
Statut éditorial
Publié
Date de publication
2012
Peer-reviewed
Oui
Volume
384
Numéro
1-2
Pages
118-127
Langue
anglais
Notes
Publication types: Comparative Study ; Journal Article
Publication Status: ppublish
Résumé
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 11weeks 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.
Mots-clé
Biotechnology/instrumentation, Biotechnology/methods, Cryopreservation, DNA/blood, DNA/genetics, Dried Blood Spot Testing/instrumentation, Dried Blood Spot Testing/methods, Gene Rearrangement, T-Lymphocyte, Humans, Leukocytes, Mononuclear, Polymerase Chain Reaction/methods, Reagent Kits, Diagnostic, Receptors, Antigen, T-Cell/genetics, Reproducibility of Results, Serum Albumin/genetics, Time Factors
Pubmed
Création de la notice
15/04/2015 8:15
Dernière modification de la notice
20/08/2019 14:38
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