A systematic analysis of PCR contamination.
In light of the strict legal scrutiny surrounding DNA typing at this time, it has become necessary to systematically address the issue of PCR contamination. To precisely define the parameters affecting PCR contamination under casework analysis conditions, PCR amplification reactions were intentionally compromised by employing sub-standard laboratory technique and by introducing secondary sources of DNA. The PCR parameters considered for potential sources of contamination include amplification set-up, amplification product handling, aerosol DNA and storage. In addition, analyst technique was evaluated by modifying or eliminating standard safeguards. Under the circumstances normally encountered during casework analysis, PCR contamination was never noted. Significantly, using the dot blot detection method, contamination was never observed when nanogram quantities of genomic DNA were mishandled or aerosolized. Contamination occurred only when amplification product was carelessly manipulated or purposefully sprayed near or directly into open tubes containing water or genomic DNA. Although standard precautions should be employed during PCR-based DNA typing, our data indicates that contamination during amplification procedures is not prevalent when detected by dot blot analysis.
Scherczinger
C A
CA
Connecticut State Police, Forensic Science Laboratory, Meriden, USA.
Ladd
C
C
Bourke
M T
MT
Adamowicz
M S
MS
Johannes
P M
PM
Scherczinger
R
R
Beesley
T
T
Lee
H C
HC
D004247:DNA / Q000032:analysis*; D016172:DNA Fingerprinting / Q000379:methods* / Q000592:standards; D004866:Equipment Contamination / Q000517:prevention & control; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity
D004247:DNA