DNA from buccal swabs recruited by mail: evaluation of storage effects on long-term stability and suitability for multiplex polymerase chain reaction genotyping.

Author(s): Freeman B, Smith N, Curtis C, Huckett L, Mill J, Craig IW

Publication: Behav Genet, 2003, Vol. 33, Page 67-72

PubMed ID: 12645823 PubMed Review Paper? No

Purpose of Paper

Conclusion of Paper

Studies

1. Study Purpose

   The purpose of this study was to evaluate the quantity and quality of DNA extracted from buccal swabs and to assess suitability for real-time qPCR analysis.

   Summary of Findings:

   The protocol outlined by the authors enables recovery of high quality DNA in quantities suitable for thousands of qPCR experiments. DNA quantity, quality, and real-time qPCR results were not altered by differences in starting concentration of DNA, room-temperature storage of buccal samples for over a year in buffer prior to processing, or frozen storage of extracted DNA for up to four years. The results were comparable to those obtained with a commercial, resin-based extraction kit (Nucleon).

   Biospecimens

   • Cell - Buccal

   Preservative Types

   • None (Fresh)

   Diagnoses:

   • Not specified

   Platform:

   Analyte	Technology Platform
   DNA	Spectrophotometry
   DNA	Electrophoresis
   DNA	Real-time qPCR

   Pre-analytical Factors:

   Classification	Pre-analytical Factor	Value(s)
   Real-time qPCR Specific	Template/input amount	5 ug 10 ug 20 ug 40 ug 80 ug
   Storage	Time at room temperature	1 week > 1 yr
   Storage	Storage duration	0 yr 4 yr
   Analyte Extraction and Purification	Analyte isolation method	Method developed by authors Commercial kit (Nucleon)

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2. Study Purpose

   The purpose of this study was to determine if the DNA extraction method reported for buccal specimens can be successfully applied to lymphocytes isolated from whole blood.

   Summary of Findings:

   Preliminary evaluation of the DNA extraction method outlined by the authors was successfully applied to lymphocytes isolated from whole blood, indicating that this extraction method may also be effective for alternative biospecimen locations. Storage of extracted DNA by both extraction methods examined generated similar multiplex real-time qPCR results.

   Biospecimens

   • Cell - White Cells

   Preservative Types

   • None (Fresh)

   Diagnoses:

   • Not specified

   Platform:

   Analyte	Technology Platform
   DNA	Real-time qPCR

   Pre-analytical Factors:

   Classification	Pre-analytical Factor	Value(s)
   Analyte Extraction and Purification	Analyte isolation method	Method developed by authors Commerical kit (Nucleon)
   Storage	Storage duration	0 yr 3-4 yr

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