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[Effect of PCR reaction volume on the accuracy of human identification tests]
Huai-gu Zhou1, Yuan Ping, Qing-wen Xu
1Shanghai Institute of Criminal Science and Technology, Shanghai 200083. hgzhou@public2.sta.net.cn
Objective:
To investigate the effect of different PCR amplification volume on the accuracy of human identification test.
Methods:
Human genome DNA samples were amplified using ABI PRISM Profiler Plus kits in 50 microliters, 25 microliters, 12.5 microliters, and 6.25 microliters reaction volume, respectively. The thermocycle parameters were the same. All PCR products were then electrophoresized on ABI PRISM 310 Genetic Analyzer, 377 DNA Sequencer, and 3100 Genetic Analyzer. Data were processed by ABI PRISM GeneScan and Genotyper software.
Results:
The less reaction volume, the more alleles losing or alleles adding observed.
Conclusion:
Non-standard volume of PCR amplification reaction should be used carefully in human identification test, especially when the sample DNA quality is not so satisfied.
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