Flexibility of GenePrintTM STR Systems Using a Low Amount of DNA
Ugo Ricci, M.L. Giovannucci Uzielli
Criminal Laboratory Department, Florence, Italy
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In our laboratory, we routinely use 7 loci of GenePrintTM STR Systems for paternity tests, linkage study and forensic application. For monoplex amplification, we employ MJ Research Thermal Cycling mod. PTC 100TM and the parameters suggested by the Technical Manual
(v. 2/96, Part # TMD004, Promega Corporation). However, when the amount of DNA is too low, or partially degraded, we sometimes verify a failure in the amplification. In these cases, the strategy suggested is to increase the volume of template, reducing the sterile water in the Master Mix reaction. This method, however, consequently increases the inhibitors of the PCR, specifically when the DNA was extracted with a non-organic procedure from stamps, cigarette butts, or contaminated samples.In these cases, we prefer these modified protocols to amplify the DNA, using 1 - 5 µL of template DNA and increasing the number of cycles.
For F13B, FESFPS and vWA: For CSF1PO, T13A01, TH01 and TPOX:
96°C 2.0 min (hot start) 96°C 2.0 min (hot start)
94°C 1.0 min 94°C 1.0 min
60°C 1.0 min 64°C 1.0 min
70°C 1.5 min 70°C 1.5 min
For 12 cycles: For 12 cycles:
90°C 1.0 min 90°C 1.0 min
60°C 1.0 min 64°C 1.0 min
70°C 1.5 min 70°C 1.5 min
For 21 or 22 cycles: For 21 or 22 cycles:
72°C 10 min 72°C 10 min
We verified flexibility of the STR System and a positive result of the amplification, without formation of undesired extra bands.
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