| Titre : | Maximizing DNA Yield for Epidemiologic Studies : No More Buffy Coats ? (2013) |
| Auteurs : | Mitchell-H GAIL ; Division of Cancer Epidemiology and Genetics, National Cancer Institute (Rockville MD, Etats-Unis) ; Tim SHEEHY ; Mark COSENTINO ; Laboratory of Biospecimen and Biorepository Research, Frederick National Laboratory for Cancer Research (Frederick MD, Etats-Unis) ; David PEE ; Norma-A DIAZ-MAYORAL ; Montserrat GARCIA-CLOSAS ; Neil-E CAPORASO ; Karen PITT ; Regina-G ZIEGLER |
| Type de document : | Article |
| Dans : | American journal of epidemiology (vol. 178, n° 7, Octobre 2013) |
| Pagination : | 1170-1176 |
| Langues: | Anglais |
| Mots-clés : | Méthodologie ; ADN ; Epidémiologie ; Cellule sanguine |
| Résumé : | [BDSP. Notice produite par INIST-CNRS tr8R0xHr. Diffusion soumise à autorisation]. Some molecular analyses require microgram quantities of DNA, yet many epidemiologic studies preserve only the buffy coat. In Frederick, Maryland, in 2010, we estimated DNA yields from 5 mL of whole blood and from equivalent amounts of all-cell-pellet (ACP) fraction, buffy coat, and residual blood cells from fresh blood (n=10 volunteers) and from both fresh and frozen blood (n=10). We extracted DNA with the OIAamp DNA Blood Midi Kit (Oiagen Sciences, Germantown, Maryland) for silica spin column capture and measured double-stranded DNA. Yields from frozen blood fractions were not statistically significantly different from those obtained from fresh fractions. ACP fractions yielded 80.6% (95% confidence interval : 66,97) of the yield of frozen whole blood and 99.3% (95% confidence interval : 86,100) of the yield of fresh blood. Frozen buffy coat and residual blood cells each yielded only half as much DNA as frozen ACP, and the yields were more variable. Assuming that DNA yield and quality from frozen ACP are stable, we recommend freezing plasma and ACP. Not only does ACP yield twice as much DNA as buffy coat but it is easier to process, and its yield is less variable from person to person. Long-term stability studies are needed. If one wishes to separate buffy coat before freezing, one should also save the residual blood cell fraction, which contains just as much DNA. |
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