Collecting and Handling Samples for Parentage and Forensics DNA‐Based Genetic Testing

David H. Bing1, Frederick R. Bieber2

1 Center for Blood Research, Boston, Massachusetts, 2 Brigham and Women's Hospital, Boston, Massachusetts
Publication Name:  Current Protocols in Human Genetics
Unit Number:  Unit 14.2
DOI:  10.1002/0471142905.hg1402s16
Online Posting Date:  May, 2001
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Abstract

The unit covers Variable Numbers of Tandem Repeats (VNTR)based paternity analysis as well as the newer methods relying on PCR to analyze sequencespecific polymorphisms and microsatellite regions. The discussion of data analysis and probability calculations has been expanded to address a number of special circumstances, such as the lack of sample from an alleged father, motherless cases, and more.

     
 
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Table of Contents

  • Basic Protocol 1: Collecting Buccal Swab Samples for DNA Testing
  • Basic Protocol 2: Collecting Blood from a Finger Prick onto FTA Paper for DNA Testing
  • Alternate Protocol 1: Procedure for Collecting Blood onto FTA Paper from Liquid EDTA‐Treated Blood
  • Commentary
  • Literature Cited
     
 
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Materials

Basic Protocol 1: Collecting Buccal Swab Samples for DNA Testing

  Materials
  • Individual to be tested
  • Powder‐free disposable gloves
  • Sterile cotton swabs (Dynarex), sterile cytology brushes, or foam oral swabs (Sage) in protective jackets
  • Evidence tape (e.g., Fitzco)
CAUTION: Do NOT touch the swab with bare hands. Wear powder‐free disposable gloves throughout the procedure.

Basic Protocol 2: Collecting Blood from a Finger Prick onto FTA Paper for DNA Testing

  Materials
  • Powder‐free disposable gloves
  • FTA filter paper (Fitzco)
  • Individual to be tested
  • Alcohol prep pads
  • Sterile gauze pads
  • Sterile lancet
  • Evidence envelope and tape (e.g., Fitzco)
CAUTION: Do NOT touch FTA paper with bare hands. Wear powder‐free disposable gloves throughout the procedure.

Alternate Protocol 1: Procedure for Collecting Blood onto FTA Paper from Liquid EDTA‐Treated Blood

  • Blood sample collected in EDTA (purple‐top) Vacutainer tubes (Becton‐Dickinson Primary Care Diagnostics)
  • Pipet tips, sterile
CAUTION: Do NOT touch FTA paper with bare hands. Wear powder‐free disposable gloves throughout the procedure.
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Figures

Videos

Literature Cited

Literature Cited
   Bever, R.A. and DeGulielmo, M.A. 1994. Development and validation of buccal swab collection method for DNA testing for paternity testing. Adv. Forensic. Haemogenet. 5:199‐201.
   Budowle, B. 1995. Guidelines for quality assurance program for DNA analysis methods. Crime Lab. Dig. 22:21‐43.
   Budowle, B., Lindsey, J.A., DeCou, J.A., Koons, B.W., Giusti, A.M., and Comey, C.T. 1995. Validation and population studies of the loci LDLR, GYPA, HBGG, D7S8, and GC (PM loci) and HLA DQα using a multiplex amplification and typing procedure. J. Forensic Sci. 40:45‐54.
   Committee on DNA Forensic Science 1996. The Evaluation of Forensic DNA Evidence. National Research Council, National Academy Press, Washington, D.C.
   DeForest, P.R., Gaensslen, R.E., and Lee, H.C. 1983. Forensic Science: An Introduction to Criminalistics. McGraw‐Hill, New York.
   FBI Laboratory 1990. Procedures for the Detection of Restriction Fragment Length Polymorphisms in Human DNA. FBI Laboratory, Washington, D.C.
   FBI Laboratory 1996. PCR‐Based Typing Protocols. FBI Laboratory, Washington, D.C.
   Hochmeister, M.N., Budowle, B., Jung, J., Borer, U.V., Comey, C.T., and Dirnhofer, R. 1991. PCR‐based typing of DNA extracted from cigarette butts. Int. J. Leg. Med. 104:229‐233.
   Holland, M., Fisher, D.L., Roby, R.K., Ruderman, J., Bryson, C., and Weedn, V. 1995. Mitochondrial DNA sequence analysis of human remains. Crime Lab. Dig. 22:109‐115.
   Kanter, F., Baird, M., Shaler, R., and Balazs, I. 1986. Analysis of restriction fragment length polymorphisms on dexoyribonucleic acid (DNA) recovered from dried blood stains. J. Forensic Sci. 31:402‐408.
   Lee, H.C., Gaensslen, R.E., Bigbee, P.D., and Kearney, J.J. 1994. Guidelines for the Collection and Preservation of DNA Evidence. U.S. Department of Justice, Federal Bureau of Investigation, Washington, D.C.
   McNally, L., Shaler, R.C., Baird, M., Balazs, I., Deforest, P., and Kobilinsky, L. 1989. Evaluation of deoxyribonucleic acid (DNA) isolated from human blood stains exposed to ultraviolet light, heat, humidity, and soil contamination. J. Forensic Sci. 34:1059‐1069.
   Tahir, M.A. and Watson, C.A. 1995. Typing of DNA HLA DQα alleles extracted from human nail material using polymerase chain reaction. J. Forensic Sci. 40:634‐636.
   Tahir, M.A., Caruso, J.F., Hamby, P.P., Sovinski, S.M., and Tahir, U.A. 1995. Restriction fragment length polymorphism (RFLP) typing of DNA extracted from nasal secretions. J. Forensic Sci. 40:459‐463.
   Wilson, M.R., Polanskey, D., Butler, J., DiZinno, J.A., Repogle, J., and Budowle, B. 1995. Extraction, PCR amplification, and sequencing of mitochondrial DNA from human hair shafts. BioTechniques. 18:662‐669.
   Word, C.J., Sawosik, T.M., and Bing, D.H. 1997. Summary of validation studies from twenty‐six forensic laboratories in the United States and Canada on the use of the AmpliType PM PCR Amplification and Typing Kit. J. Forensic Sci. 42:39‐48.
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