Comparison of three DNA extraction methods on bone and blood stains up to 43 years old and amplification of three different gene sequences.

Author(s): Cattaneo C, Craig OE, James NT, Sokol RJ

Publication: J Forensic Sci, 1997, Vol. 42, Page 1126-35

PubMed ID: 9397557 PubMed Review Paper? No

Purpose of Paper

Conclusion of Paper

Studies

1. Study Purpose

   The purpose of this study was to determine the effects of storage, extraction method, targeted nucleic acid and amplicon size on the extraction of amplifiable DNA from bone and blood specimens. Blood smears were dried and stored at room temperature for 0-26 years. Bone specimens were obtained fresh, stored for 3-6 years at 4 degrees C, or interred for up to 40 years then stored at ambient temperatures for 5 years followed by storage at -80 degrees C.

   Summary of Findings:

   Amplifiable DNA was obtained more often from dried-blood specimens than bone specimens in which a significant decline in amplifiability was noted in the first 10 years of storage. DNA extracted using the glass milk method had the highest rate of amplification success (65%) followed by DNA extracted with sodium acetate (55%) and magnetic beads (37%). Sodium acetate extraction yielded the largest quantity of DNA, but resulted in significant PCR inhibition (24%) in contrast to the low PCR inhibition observed when extraction was performed using glass milk (6%) or magnetic beads (11%). Mitochondrial DNA was most often successfully amplified (84%). No differences were observed between HLA DPB1 (39%) and amelogenin (35%) amplification despite the shorter amplicon length of amelogenin (106 bp versus 327 bp).

   Biospecimens

   • Tissue - Bone
   • Bodily Fluid - Blood

   Preservative Types

   • None (Fresh)
   • Frozen
   • Other Preservative

   Diagnoses:

   • Autopsy
   • Not specified

   Platform:

   Analyte	Technology Platform
   DNA	PCR
   DNA	Spectrophotometry

   Pre-analytical Factors:

   Classification	Pre-analytical Factor	Value(s)
   Biospecimen Acquisition	Biospecimen location	Bone Blood
   PCR Specific	Targeted nucleic acid	HLA DPB1 Amelogenin Mitochondrial
   PCR Specific	Length of gene fragment	106/112 bp 327 bp
   Analyte Extraction and Purification	Analyte isolation method	Sodium acetate Magnetic beads Glass milk
   Storage	Storage duration	0 years 3-6 years 13-43 years
   Biospecimen Preservation	Type of fixation/preservation	Frozen None (fresh) Refrigeration Air-dried

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