[Effect of PCR reaction volume on the accuracy of human identification tests].
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.


CONCLUSIONS
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.
Zhou
Huai-gu
HG
Shanghai Institute of Criminal Science and Technology, Shanghai 200083. hgzhou@public2.sta.net.cn
Ping
Yuan
Y
Xu
Qing-wen
QW
D000483:Alleles; D001801:Blood Stains; D004247:DNA / Q000032:analysis* / Q000235:genetics / Q000302:isolation & purification; D004586:Electrophoresis; D005554:Forensic Medicine; D006801:Humans; D019656:Loss of Heterozygosity; D016133:Polymerase Chain Reaction / Q000379:methods; D012680:Sensitivity and Specificity; D013056:Spectrophotometry, Ultraviolet
D004247:DNA