Maximizing DNA Yield for Epidemiologic Studies: No More Buffy Coats?

Gail, Mitchell H.; Sheehy, Tim; Cosentino, Mark; Pee, David; Diaz-Mayoral, Norma A.; Garcia-Closas, Montserrat; Caporaso, Neil E.; Pitt, Karen; Ziegler, Regina G.
October 2013
American Journal of Epidemiology;Oct2013, Vol. 178 Issue 7, p1170
Academic Journal
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 QIAamp DNA Blood Midi Kit (Qiagen 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.


Related Articles

  • Toward a patient-based paradigm for blood transfusion. Farrugia, Albert; Vamvakas, Eleftherios // Journal of Blood Medicine;2014, Vol. 5, p5 

    The current "manufacturing paradigm" of transfusion practice has detached transfusion from the clinical environment. As an example, fresh whole blood in large-volume hemorrhage may be superior to whole blood reconstituted from multiple components. Multicomponent apheresis can overcome logistical...

  • Blood drive Tuesday.  // Clarendon Enterprise (TX);11/15/2007, Vol. 18 Issue 48, p4 

    The article reports that Coffee Memorial Blood Center will be hosting a blood drive on November 20, 2007 at the Bairfield Activity Center in Clarendon, Texas.

  • All types of blood donors needed.  // Consumer Reports on Health;May2000, Vol. 12 Issue 5, p2 

    Discusses the need for blood donors in the United States and the health benefits of donating blood.

  • Colonial Ridge lists day for blood drive Jan. 28.  // Breckenridge American;1/16/2008, Vol. 109, p2 

    The article calls for blood donors for the blood drive of the Colonial Ridge Health and Rehab Center's on January 28, 2008 in Breckenridge.

  • ERRATUM.  // American Journal of Public Health;Mar2014, Vol. 104 Issue 3, pe12 

    A correction notice is presented for the article "Studies With Staggered Starts: Multiple Baseline Designs and Group-Randomized Trials" by D. A. Rhoda, D. M. Murray and R. R. Andridge et al., which appeared in a 2011 issue.

  • The Effects of Rate on Single-Word Reading Assessment. Carter, Matthew D.; Walker, Marianna M.; O’Brien, Kevin // American Journal of Speech-Language Pathology;Feb2015, Vol. 24 Issue 1, p13 

    Purpose: The purpose of this study was to examine the effects of introducing rapid stimulus presentation durations while limiting response durations on the decoding profiles of college students with reading disorders. Method: Eighteen college-aged individuals with typical reading abilities and...

  • Israelis destroy Ethiopian blood donations. Siegel-Itzkovich, Judy // BMJ: British Medical Journal (International Edition);2/10/96, Vol. 312 Issue 7027, p331 

    Reports that public outcry has greeted the discovery that for years most of the blood donated by Ethiopian Jewish immigrants to Israel's blood collection service has been thrown out unused because of the high risk of HIV in the community. Apology of Prime Minister Shimon Peres.

  • Blood drive Friday in Whitney.  // Lakelander (Whitney, TX);8/12/2009, Vol. 23 Issue 32, p1 

    The article reports on the community blood drive campaign hosted by Brookshire Development with Carter BloodCare on August 14, 2009 in Whitney, Texas.

  • Good Samaritan Hospital seeks to increase blood donors in '99.  // Hudson Valley Business Journal;03/29/99, Vol. 9 Issue 24, p22 

    Reports on the Good Samaritan Hospital's receipt of community and corporate support for its volunteer blood donor program in Rockland County, New York. Plans of the hospital to increase its number of donors; Sources of blood donations; Personal benefits of donating blood.


Read the Article


Sorry, but this item is not currently available from your library.

Try another library?
Sign out of this library

Other Topics