Long-term Storage and Recovery of Buccal Cell DNA from Treated Cards

Economical methods for collecting and storing high-quality DNA are needed for large population-based molecular epidemiology studies. Buccal cell DNA collected via saliva and stored on treated filter paper cards could be an attractive method, but modest DNA yields and the potential for reduced recove...

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Bibliographic Details
Published in:Cancer epidemiology, biomarkers & prevention biomarkers & prevention, 2006-02, Vol.15 (2), p.385-388
Main Authors: Sigurdson, Alice J, Ha, Mina, Cosentino, Mark, Franklin, Tracie, Haque, Kashif A, Qi, Ying, Glaser, Cynthia, Reid, Yvonne, Vaught, Jim B, Bergen, Andrew W
Format: Article
Language:English
Subjects:
archival samples
Biological and medical sciences
biospecimen storage
buccal cells
DNA - analysis
DNA collection
Epidemiology
Humans
Medical sciences
methods
Mouth Mucosa - cytology
Nucleic Acid Amplification Techniques
Polymerase Chain Reaction
Specimen Handling - economics
Specimen Handling - instrumentation
Specimen Handling - methods
Tumors
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Summary:Economical methods for collecting and storing high-quality DNA are needed for large population-based molecular epidemiology studies. Buccal cell DNA collected via saliva and stored on treated filter paper cards could be an attractive method, but modest DNA yields and the potential for reduced recovery of DNA over time were unresolved impediments. Consequently, buccal cell DNA collection via oral mouthwash rinsing became the method of choice in epidemiologic studies. However, the amount of genomic DNA (gDNA) required for genotyping continues to decrease, and reliable whole genome amplification (WGA) methods further reduced the mass of gDNA needed for WGA to 10 ng, diminishing the obstacle of low DNA yields from cards. However, concerns about yield and DNA quality over time remained. We located and analyzed 42 buccal cell saliva samples collected and stored on treated cards for 7 years at room temperature, −20°C, and −80°C. We recovered DNA from the treated cards, estimated the concentration by a human-specific quantitative real-time PCR assay, and evaluated the quality by PCR amplification of 268-, 536-, and 989-bp fragments of the β-globin gene and by AmpF l STR Identifiler assay analysis. Most DNA yields per 3-mm punch were
ISSN:1055-9965
1538-7755
DOI:10.1158/1055-9965.EPI-05-0662