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DNA Amplification for Forensic Analysts

PowerPlex® (Promega GenePrint®)

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Promega was the first manufacturer to produce kits for STR testing. Although these kits were initially offered as monoplexes, they were instrumental in setting the foundation for forensic STR DNA analysis. Promega went on to produce several multiplexes for STR DNA analysis. One of the first multiplex amplification kits offered by Promega was the GenePrint® CTTv Multiplex amplification, which amplified CSF1PO, TPOX, THO1, and vWA using a blue fluorescein (FL) dye. In addition, Promega produced both the GenePrint® FFFL Multiplex kit, which amplified F13A1, FES/FPS, F13B, and LPL, and the GenePrint® GammaSTR Multiplex kit, which amplified D16S539, D13S317, D7S820, and D5S818. Both of these kits also used the blue FL dye. When used together these kits provided the most powerful combination of multiplexes offered for forensic STR DNA analysis at that time.

As with Profiler and Cofiler, the combination of these two kits (GenePrint® PowerPlex™ 1.1 System and GenePrint® PowerPlex™ 2.1 System ) provides laboratories with a means to meet the national requirements for analyzing the 13 CODIS core loci. They incorporate a quality control measure by ensuring concordance of the repeated loci. The first kit was PowerPlex® 1.1, for Hitachi FMBIO users, or the PowerPlex® 1.2 System, for ABI 377, 310 and 3100 users. The two dye kits amplified eight STR loci and amelogenin in the following configuration:

  • D5S818, D13S317, D7S820, and D16S539 labeled with a blue (FL) dye
  • vWA , THO1, TPOX, and CSF1PO labeled with a yellow carboxy-tetramethylrhodamide (TMR) dye.09, 10

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Later Promega introduced the PowerPlex® 2.1 System, which is designed for Hitachi FMBIO users. The two dye kit amplified nine STR loci and amelogenin in the following configuration:

  • D3S1358, THO1, D21S11, D18S51, and Penta E labeled with a blue FL dye.
  • Amelogenin, vWA, D8S1179, TPOX, and FGA labeled with a yellow TMR dye

As with Profiler and Cofiler, combining these two kits provided laboratories with a means to meet the national requirements for analyzing the 13 CODIS core loci and incorporate a quality control measure by ensuring concordance of the repeated loci (THO1, vWA TPOX, and amelogenin). An additional locus, Penta E, was added to this set providing for increased discrimination power.11

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Promega was the first commercial manufacturer in the country to offer a STR multiplex that typed all of the CODIS core loci with a single amplification. The kit, PowerPlex® 16, is a three-dye kit that amplifies 15 STR loci and amelogenin in the following configuration: 

  • D3S1358, THO1, D21S11, D18S51, and Penta E labeled with a blue FL dye
  • D5S818, D13S317, D7S820, D16S539, CSF1PO, and Penta D labeled with a green 6-carboxy-4',5'-dichloro-2',7'-dimethoxy-fluorescein (JOE) dye
  • Amelogenin, vWA, D8S1179, TPOX, and FGA labeled with a yellow TMR dye

In addition to the core loci, PowerPlex® 16 also amplifies two penta-nucleotide repeats for increased discrimination power.12

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Promega was the first commercial manufacturer to produce a Y-STR kit that amplified all of the core Y-STR loci, as outlined by SWGDAM. The PowerPlex® Y amplification kit is a three-dye chemistry kit that amplifies 12 Y-STR loci in the following configuration:

  • DYS391, DYS389I, DYS439, and DYS389II are labeled with a blue FL dye
  • DYS438, DYS437, DYS19, and DYS392 are labeled with a green JOE dye
  • DYS393, DYS390, andDYS385 A/B are labeled with a yellow TMR dye
  • The PowerPlex® Y kit amplifies all of the required core Y-STR loci in addition to DYS437.

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