Cynthia J. Sprecher, Katherine A. Micka, Ann M. Lins and James W. Schumm
Promega Corporation, 2800 Woods Hollow Road, Madison, WI
Short Tandem Repeats (STRs) are polymorphic PCR-based systems useful in human identification. High throughput analyses of STRs is achieved with the GenePrint™ STR Multiplex Systems which allow for amplification of up to 4 STR loci simultaneously. The systems, CSF1PO-TPOX-TH01 (CTT) and F13A01-FESFPS-vWA (FFv) for silver detection, CSF1PO-TPOX-TH01-vWA (CTTv) and F13A01-FESFPS-F13B-LPL (FFFL) for fluorescent detection, were originally developed for use on the Perkin Elmer Model 480 Thermal Cycler.
Differences in the features of the Perkin Elmer Model 9600 Thermal Cycler can cause performance differences with these multiplex systems. Sometimes protocol modification for use with the Perkin Elmer Model 9600 Thermal Cycler improves performance avoiding changes in amplification sensitivity or product yield. Protocol differences are less important for the CTT multiplex, and most obvious with the FFv system. Well defined protocol options which provide excellent results for each STR multiplex using each thermal cycler will be presented.
In addition, we have discovered that only moderate changes in amplification sensitivity and product yield are observed with some GenePrint™ STR Multiplex Systems when abbreviated protocols are used. Once again, different systems reveal different responses to these effects, with the CTTv quadriplex being most sensitive to shortening of protocol times, and the others showing very little effect.
Comparison of several amplification protocols with each GenePrint™ STR Multiplex System will be demonstrated.
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