Biochemical Genetics, VoL 3i, Nos. 7/8, I993
DNA Isolation by a Rapid Method from Human
Blood Samples: Effects of MgCI2, EDTA, Storage
Time, and Temperature on DNA Yield and Quality
Dcbomoy K. LahirP ,2 and Bill SchnabeP
Received 5 Mar, 1993 Final 22 June 1993
The isolation of DNA from whole blood by a modified rapid method (RM) was
tested using various detergents and buffer conditions. Extraction of DNA with
either NP-40 or Triton X-IO0 gave a high yield of undegraded DNA in less than
an hour. The concentration of magnesium ion in the buffers was critical to
obtaining intact, high molecular weight (HMW) DNA. Greater than 10 mM
MgCl2 led to degradation. Addition of EDTA to the buffer inhibits this degrada-
tion. Preparation of DNA from blood stored at room temperature or incubated at
37~C for 24 hr resulted in the same amount and quality of DNA as from samples
frozen at - 70°C. DNA from blood samples that had undergone more than four
freeze-thaw cycles was found to be pal¢ially degraded. The modified RM can be
applied to extract DNA from as little as 10 ixl of blood (340 ng of DNA) and
from dried blood samples. DNA samples remained intact and undegraded for
longer times when DNA was dissolved in higher concentrations of EDTA.
KEY WORDS: high molecular weight DNA; MgClz; inte~ity of DNA; rapid method; DNA
banking.
INTRODUCTION
The application of genetics to study human disease (Anderson, 1992; Miller,
1992) and to analyze gene function in vivo depends upon the quality of DNA
samples. Intact and good-quality DNA is also essential for screening DNA
This work was supported by grants from the Indiana Department of Mental Health and PHS
RO1 AG10297.
i Laboratory of Molecular Neurogenetics, Institute of Psychiatric Research, Department of
Psychiatry, Indiana University School of Medicine, Indianapolis, Indiana 46202-4887.
2 To whom correspondence should be addressed at Laboratory of Molecular Neurogenetics,
Institute of Psychiatric Research, Indiana University School of Medicine, 791 Union Drive,
Indianapolis, Indiana 46202-4887.
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0006-2928/93/0800-0321507.00/0 ©i993Plenum Publishing Corporation