From Silver to Multicolor Fluorescence: STR Multiplex Systems
Cynthia J. Sprecher, Katherine A. Micka, Ann M. Lins, Dawn R. Rabbach, Jennifer A.
Taylor, Jeffrey W. Bacher, and James W. Schumm
Promega Corporation, 2800 Woods Hollow Road, Madison, WI 53711 USA.
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We have developed STR multiplex systems for a variety of detection methods including silver stain as well as single-color and multi-color fluorescent detection. For each of the STR loci in these multiplexes, we have developed allelic ladders, which have been characterized by sequence analysis.
Multiplex systems, for silver stain detection include the GenePrint
Ô STR Multiplex-CSF1PO, TPOX, TH01 (CTT) and the GenePrintÔ STR Multiplex-F13A01, FESFPS, vWA (FFv) which allow for co-amplification of three STR loci. A third multiplex being developed for silver detection is the SilverSTRÔ III Multiplex includes the three STR loci, D16S539, D7S820, and D13S317.We have also developed three multiplex systems for use with single color fluorescent detection. The GenePrintTM Fluorescent STR Systems F13A01, FESFPS, F13B, LPL (FFFL), CSF1PO, TPOX, TH01, vWA (CTTv), and GammaSTR
Ô (D16S539, D7S820, D13S317, D5S818) allow simultaneous amplification, detection, and analysis of 4 polymorphic STR loci.The GenePrintTM PowerPlexTM System has been developed to combine eight polymorphic STR loci (CSF1PO, TPOX, TH01, vWA, D16S539, D7S820, D13S317, D5S818) in a single amplification. The loci are detected with a two-color fluorescent system.
In multi-color detection, a third fluorescent dye can be used for an internal size standard. This internal size standard can be used for determining the size of alleles reducing the number of allelic ladder lanes required. Allelic ladders are still needed as the sizes of alleles calculated using an internal size standard vary with the gel format and with the size standard used.
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