Logistics and quality control for DNA sampling in large multicenter studies

Summary.  To study associations between genetic variation and disease, large bio‐banks need to be created in multicenter studies. Therefore, we studied the effects of storage time and temperature on DNA quality and quantity in a simulation experiment with storage up to 28 days frozen, at 4 °C and at room temperature. In the simulation experiment, the conditions did not influence the amount or quality of DNA to an unsatisfactory level. However, the amount of extracted DNA was decreased in frozen samples and in samples that were stored for > 7 days at room temperature. In a sample of patients from 24 countries of the EUROPA trial obtained by mail with transport times up to 1 month DNA yield and quality were adequate. From these results we conclude that transport of non‐frozen blood by ordinary mail is usable and practical for DNA isolation for polymerase chain reaction in clinical and epidemiological studies.

[1]  B. Korf,et al.  Preliminary phenotypic map of chromosome 4p16 based on 4p deletions. , 1995, American journal of medical genetics.

[2]  Sophie Visvikis,et al.  DNA Extraction and Stability for Epidemiological Studies , 1998, Clinical chemistry and laboratory medicine.

[3]  D. Lahiri,et al.  DNA isolation by a rapid method from human blood samples: Effects of MgCl2, EDTA, storage time, and temperature on DNA yield and quality , 1993, Biochemical Genetics.

[4]  R. Sepp,et al.  Rapid techniques for DNA extraction from routinely processed archival tissue for use in PCR. , 1994, Journal of clinical pathology.

[5]  E. Keystone,et al.  Analysis of FcgammaRII gene polymorphisms in Wegener's granulomatosis. , 1997, Experimental and clinical immunogenetics.

[6]  D. H. Farkas,et al.  Specimen collection and storage for diagnostic molecular pathology investigation. , 1996, Archives of pathology & laboratory medicine.

[7]  J. Brockmöller,et al.  Arylamine N-acetyltransferase (NAT2) mutations and their allelic linkage in unrelated Caucasian individuals: correlation with phenotypic activity. , 1995, American journal of human genetics.

[8]  J. Jespersen,et al.  Internal Quality Control of PCR-based Genotyping Methods in Research Studies and Patient Diagnostics , 2002, Thrombosis and Haemostasis.

[9]  J. Kersey,et al.  A modified method of DNA extraction from peripheral blood and bone marrow specimens , 1988, American journal of hematology.

[10]  J. Medrano,et al.  Effects of blood storage time and temperature on DNA yield and quality. , 1993, BioTechniques.

[11]  N. N. Brown,et al.  Routine use of hair root or buccal swab specimens for PCR analysis: advantages over using blood. , 1992, Clinica chimica acta; international journal of clinical chemistry.

[12]  J. Sambrook,et al.  Molecular Cloning: A Laboratory Manual , 2001 .

[13]  Shirley A. Miller,et al.  A simple salting out procedure for extracting DNA from human nucleated cells. , 1988, Nucleic acids research.

[14]  L. Madisen,et al.  DNA banking: the effects of storage of blood and isolated DNA on the integrity of DNA. , 1987, American journal of medical genetics.

[15]  A. Lew,et al.  PCR based diagnosis in the presence of 8% (v/v) blood. , 1991, Nucleic acids research.

[16]  K. Fox,et al.  The EUROPA Trial: Design, Baseline Demography and Status of the Substudies , 2001, Cardiovascular Drugs and Therapy.

[17]  E. Keystone,et al.  Analysis of Fc?RII Gene Polymorphisms in Wegeners Granulomatosis , 1997 .

[18]  L. Tobler,et al.  Impact of specimen handling and storage on detection of hepatitis C virus RNA , 1992, Transfusion.

[19]  M. Neumaier,et al.  Fundamentals of quality assessment of molecular amplification methods in clinical diagnostics. International Federation of Clinical Chemistry Scientific Division Committee on Molecular Biology Techniques. , 1998, Clinical chemistry.