1. Manuals
  2. Brands
  3. Promega Manuals
  4. Measuring Instruments
  5. PowerPlex ESI 17 Fast System
  6. Technical manual

Promega PowerPlex ESI 17 Fast System Technical Manual

Applied biosystems genetic analyzers
Hide thumbs
Table of Contents

Advertisement

Quick Links

Download this manual
T E C H N I C A L M A N U A L
PowerPlex
for Use on the
Applied Biosystems
Genetic Analyzers
Instructions for Use of Products
DC1720 and DC1721
ESI 17 Fast System
®
®
Revised 10/23
TMD041

Advertisement

Table of Contents
loading
Need help?

Need help?

Do you have a question about the PowerPlex ESI 17 Fast System and is the answer not in the manual?

Questions and answers

Related Manuals for Promega PowerPlex ESI 17 Fast System

Summary of Contents for Promega PowerPlex ESI 17 Fast System

  • Page 1 T E C H N I C A L M A N U A L PowerPlex ESI 17 Fast System ® for Use on the Applied Biosystems ® Genetic Analyzers Instructions for Use of Products DC1720 and DC1721 Revised 10/23 TMD041...

  • Page 2: Table Of Contents

    ID Software, Version 3.2 ....51 ® 6.K. Controls ............................. 54 6.L. Results ............................54 Promega Corporation · 2800 Woods Hollow Road · Madison, WI 53711-5399 USA · Toll Free in USA 800-356-9526 · 608-274-4330 · Fax 608-277-2516 · www.promega.com TMD041 Revised 10/23...
  • Page 3 9.E. Composition of Buffers and Solutions ..................... 90 9.F. Related Products.......................... 91 9.G. Summary of Changes ........................91 Promega Corporation · 2800 Woods Hollow Road · Madison, WI 53711-5399 USA · Toll Free in USA 800-356-9526 · 608-274-4330 · Fax 608-277-2516 TMD041 · Revised 10/23 www.promega.com...

  • Page 4: Description

    Information about other Promega fluorescent STR systems is available upon request from Promega or online at: www.promega.com Promega Corporation · 2800 Woods Hollow Road · Madison, WI 53711-5399 USA · Toll Free in USA 800-356-9526 · 608-274-4330 · Fax 608-277-2516 ·...

  • Page 5: Product Components And Storage Conditions

    ESI 17 Fast System components may be stored for up to 1 year at +2°C to +10°C without ® loss of activity. Promega Corporation · 2800 Woods Hollow Road · Madison, WI 53711-5399 USA · Toll Free in USA 800-356-9526 · 608-274-4330 · Fax 608-277-2516 TMD041 · Revised 10/23 www.promega.com...

  • Page 6: Before You Begin

    (i.e., no template) to detect reagent contamination. We highly recommend the use of gloves and aerosol-resistant pipette tips. Promega Corporation · 2800 Woods Hollow Road · Madison, WI 53711-5399 USA · Toll Free in USA 800-356-9526 · 608-274-4330 · Fax 608-277-2516 ·...

  • Page 7: Spectral Calibration

    Meticulous care must be taken to ensure successful amplification. A guide to amplification troubleshooting is provided in Section 7. Promega Corporation · 2800 Woods Hollow Road · Madison, WI 53711-5399 USA · Toll Free in USA 800-356-9526 · 608-274-4330 · Fax 608-277-2516 TMD041 · Revised 10/23...

  • Page 8: Amplification Of Extracted Dna In A 25Μl Reaction Volume

    Use a clean plate for reaction assembly, and label it appropriately. Alternatively, determine the number of clean, 0.2ml reaction tubes required, and label them appropriately. Promega Corporation · 2800 Woods Hollow Road · Madison, WI 53711-5399 USA · Toll Free in USA 800-356-9526 · 608-274-4330 · Fax 608-277-2516 ·...

  • Page 9: Amplification Of Extracted Dna In A 25Μl Reaction Volume

    DNA template used in your laboratory should be based on the results of your internal validation and may be different. Promega Corporation · 2800 Woods Hollow Road · Madison, WI 53711-5399 USA · Toll Free in USA 800-356-9526 · 608-274-4330 · Fax 608-277-2516 TMD041 · Revised 10/23...
  • Page 10 –20°C protected from light. Note: Long-term storage of amplified samples at 4°C or higher may produce artifacts. Promega Corporation · 2800 Woods Hollow Road · Madison, WI 53711-5399 USA · Toll Free in USA 800-356-9526 · 608-274-4330 · Fax 608-277-2516 ·...

  • Page 11: Direct Amplification Of Dna From Storage Card Punches In A 25Μl Reaction Volume

    1.2mm sample disk. Use the plunger to eject the disk into the appropriate well of a reaction plate. Promega Corporation · 2800 Woods Hollow Road · Madison, WI 53711-5399 USA · Toll Free in USA 800-356-9526 · 608-274-4330 · Fax 608-277-2516 TMD041 · Revised 10/23...
  • Page 12 Use a clean plate for reaction assembly, and label it appropriately. Alternatively, determine the number of clean, 0.2ml reaction tubes required, and label them appropriately. Promega Corporation · 2800 Woods Hollow Road · Madison, WI 53711-5399 USA · Toll Free in USA 800-356-9526 · 608-274-4330 · Fax 608-277-2516 ·...
  • Page 13 For lytic storage cards, add one or two 1.2mm punches from a card containing a buccal sample or one 1.2mm punch from a card containing whole blood to the appropriate wells of the reaction plate. For nonlytic storage card punches, add the PCR amplification mix to the PunchSolution™ Reagent-treated punches. Note: It also is acceptable to add the lytic storage card punch first, and then add the PCR amplification mix. For the positive amplification control, vortex the tube of 2800M Control DNA then add 1μl (10ng) to a reaction well containing 25μl of PCR amplification mix. Notes: Do not include blank storage card punches in the positive control reactions. Optimization of the amount of 2800M Control DNA may be required, depending on thermal cycling conditions and laboratory preferences. Promega Corporation · 2800 Woods Hollow Road · Madison, WI 53711-5399 USA · Toll Free in USA 800-356-9526 · 608-274-4330 · Fax 608-277-2516 TMD041 · Revised 10/23 www.promega.com...
  • Page 14 –20°C protected from light. Note: Long-term storage of amplified samples at 4°C or higher may produce artifacts. Promega Corporation · 2800 Woods Hollow Road · Madison, WI 53711-5399 USA · Toll Free in USA 800-356-9526 · 608-274-4330 · Fax 608-277-2516 ·...

  • Page 15: Direct Amplification Of Dna From Swabs In A 25Μl Reaction Volume

    Do not centrifuge the 10X Primer Pair Mix or 5X Master Mix after vortexing, as this may cause the reagents to be concentrated at the bottom of the tube. Promega Corporation · 2800 Woods Hollow Road · Madison, WI 53711-5399 USA · Toll Free in USA 800-356-9526 · 608-274-4330 · Fax 608-277-2516 TMD041 · Revised 10/23...
  • Page 16 Note: Optimization of the amount of 2800M Control DNA may be required depending on thermal cycling conditions and laboratory preferences. Promega Corporation · 2800 Woods Hollow Road · Madison, WI 53711-5399 USA · Toll Free in USA 800-356-9526 · 608-274-4330 · Fax 608-277-2516 ·...
  • Page 17 Note: Long-term storage of amplified samples at 4°C or higher may produce artifacts. Promega Corporation · 2800 Woods Hollow Road · Madison, WI 53711-5399 USA · Toll Free in USA 800-356-9526 · 608-274-4330 · Fax 608-277-2516 TMD041 · Revised 10/23...

  • Page 18: Instrument Setup And Sample Preparation

    Always wear gloves and safety glasses when working with formamide. Promega Corporation · 2800 Woods Hollow Road · Madison, WI 53711-5399 USA · Toll Free in USA 800-356-9526 · 608-274-4330 · Fax 608-277-2516 ·...
  • Page 19 Centrifuge plate briefly to remove air bubbles from the wells. Denature samples at 95°C for 3 minutes, and then immediately chill on crushed ice, a freezer plate block or in an ice-water bath for 3 minutes. Denature samples just prior to loading the instrument. Promega Corporation · 2800 Woods Hollow Road · Madison, WI 53711-5399 USA · Toll Free in USA 800-356-9526 · 608-274-4330 · Fax 608-277-2516 TMD041 · Revised 10/23 www.promega.com...
  • Page 20 Set the oven temperature to 60°C, and then select Start Pre-heat. When the Oven Temperature and Detection Cell Temperature turn green, you can proceed with the first injection. Figure 1. The Dashboard. Promega Corporation · 2800 Woods Hollow Road · Madison, WI 53711-5399 USA · Toll Free in USA 800-356-9526 · 608-274-4330 · Fax 608-277-2516 · www.promega.com...
  • Page 21 Applied Biosystems 3500/3500xL Genetic Analyzer User Guide to edit a library entry. Assign a descriptive protocol name. Note: For more detailed information refer to the Applied Biosystems 3500/3500xL Genetic Analyzer User Guide. Promega Corporation · 2800 Woods Hollow Road · Madison, WI 53711-5399 USA · Toll Free in USA 800-356-9526 · 608-274-4330 · Fax 608-277-2516 TMD041 · Revised 10/23 www.promega.com...
  • Page 22 Run Time 1,210 seconds Injection time may be modified to increase or decrease peak heights. Promega Corporation · 2800 Woods Hollow Road · Madison, WI 53711-5399 USA · Toll Free in USA 800-356-9526 · 608-274-4330 · Fax 608-277-2516 · www.promega.com TMD041...
  • Page 23 325, 350, 375, 400, 425, 450, 475 and 500 bases. See Figure 3. Figure 3. The Create New Size Standard window. Promega Corporation · 2800 Woods Hollow Road · Madison, WI 53711-5399 USA · Toll Free in USA 800-356-9526 · 608-274-4330 · Fax 608-277-2516 TMD041 · Revised 10/23...
  • Page 24 3500 or 3500xL Genetic Analyzer. Figure 4 shows one option for these ® settings. Figure 4. The ‘Create New QC Protocol’ window. Promega Corporation · 2800 Woods Hollow Road · Madison, WI 53711-5399 USA · Toll Free in USA 800-356-9526 · 608-274-4330 · Fax 608-277-2516 · www.promega.com TMD041...
  • Page 25 Note: If autoanalysis of a sample is desired, refer to the instrument user’s manual for instructions. Figure 5. The ‘Create New Assay’ window. Promega Corporation · 2800 Woods Hollow Road · Madison, WI 53711-5399 USA · Toll Free in USA 800-356-9526 · 608-274-4330 · Fax 608-277-2516 TMD041 · Revised 10/23...
  • Page 26 Select the File Name Attributes according to laboratory practices, and save with a descriptive name. Figure 6. The ‘Create New File Name Convention’ window. Promega Corporation · 2800 Woods Hollow Road · Madison, WI 53711-5399 USA · Toll Free in USA 800-356-9526 · 608-274-4330 · Fax 608-277-2516 ·...
  • Page 27 5.A. Detection of Amplified Fragments Using the Applied Biosystems 3500 or 3500xL Genetic Analyzer (continued) ® 2.f. To create a new Results Group (Figure 7), navigate to the Library. Select Results Group, and then select Create. Alternatively, a previously created Results Group can be used. Select the Results Group Attributes according to laboratory practices. Save with a descriptive name. Figure 7. The ‘Create New Results Group’ window. Promega Corporation · 2800 Woods Hollow Road · Madison, WI 53711-5399 USA · Toll Free in USA 800-356-9526 · 608-274-4330 · Fax 608-277-2516 TMD041 · Revised 10/23 www.promega.com...
  • Page 28 Assign a descriptive plate name. Select the plate type HID from the drop-down menu (Figure 8). Figure 8. Defining plate properties. Select Assign Plate Contents (Figure 9). Figure 9. Assigning plate contents. Promega Corporation · 2800 Woods Hollow Road · Madison, WI 53711-5399 USA · Toll Free in USA 800-356-9526 · 608-274-4330 · Fax 608-277-2516 · www.promega.com TMD041...
  • Page 29 In the ‘Run Information’ window (Figure 10), assign a Run Name. Select Start Run (not shown). Note: Each injection will take approximately 40 minutes. Figure 10. Assigning a run name. Promega Corporation · 2800 Woods Hollow Road · Madison, WI 53711-5399 USA · Toll Free in USA 800-356-9526 · 608-274-4330 · Fax 608-277-2516 TMD041 · Revised 10/23 www.promega.com...

  • Page 30: Detection Of Amplified Fragments Using The Applied Biosystems ® 3130 Or 3130Xl Genetic Analyzer With Data Collection Software, Version 3.0 Or 4.0

    Vortex for 10–15 seconds to mix. Pipet 10µl of formamide/internal lane standard mix into each well. Promega Corporation · 2800 Woods Hollow Road · Madison, WI 53711-5399 USA · Toll Free in USA 800-356-9526 · 608-274-4330 · Fax 608-277-2516 ·...
  • Page 31 Note: If autoanalysis of sample data is desired, refer to the instrument user’s manual for instructions. Promega Corporation · 2800 Woods Hollow Road · Madison, WI 53711-5399 USA · Toll Free in USA 800-356-9526 · 608-274-4330 · Fax 608-277-2516 TMD041 · Revised 10/23...
  • Page 32 Monitor electrophoresis by observing the run, view, array or capillaries viewer window in the data collection software. Each injection will take approximately 40 minutes. Promega Corporation · 2800 Woods Hollow Road · Madison, WI 53711-5399 USA · Toll Free in USA 800-356-9526 · 608-274-4330 · Fax 608-277-2516 ·...

  • Page 33: Data Analysis

    Select File, and then Import Marker Stutter. A warning box will appear asking if you want to overwrite current values. Select Yes. Promega Corporation · 2800 Woods Hollow Road · Madison, WI 53711-5399 USA · Toll Free in USA 800-356-9526 · 608-274-4330 · Fax 608-277-2516 TMD041 · Revised 10/23...

  • Page 34: Importing The Wen Ils 500 Ess Size Standard Into Genemapper ® Id-X Software

    Group. This allows access for all users of the software. Other security groups may be used. Promega Corporation · 2800 Woods Hollow Road · Madison, WI 53711-5399 USA · Toll Free in USA 800-356-9526 · 608-274-4330 · Fax 608-277-2516 ·...
  • Page 35 300, 325, 350, 375, 400, 425, 450, 475 and 500 bases). See Section 9.B, Figure 23. Select OK. Promega Corporation · 2800 Woods Hollow Road · Madison, WI 53711-5399 USA · Toll Free in USA 800-356-9526 · 608-274-4330 · Fax 608-277-2516 TMD041 · Revised 10/23...

  • Page 36: Creating A Casework Analysis Method With Genemapper ® Id-X Software

    Enter a descriptive name for the analysis method, such as “PowerPlex ESI 17 Fast”. Select the ‘Allele’ tab (Figure 12). Promega Corporation · 2800 Woods Hollow Road · Madison, WI 53711-5399 USA · Toll Free in USA 800-356-9526 · 608-274-4330 · Fax 608-277-2516 ·...
  • Page 37 ® You may need to optimize these settings. In-house validation should be performed. Promega Corporation · 2800 Woods Hollow Road · Madison, WI 53711-5399 USA · Toll Free in USA 800-356-9526 · 608-274-4330 · Fax 608-277-2516 TMD041 · Revised 10/23...
  • Page 38 Notes: Select full range or partial range for the analysis range. When using a partial range, choose an appropriate analysis range based on your data. Choose a start point after the primer peak and just before the first defined internal lane standard peak to help ensure proper sizing of the internal lane standard. The peak amplitude thresholds are the minimum peak heights at which the software will call a peak. Individual laboratories should determine their peak amplitude thresholds from internal validation studies. The normalization box can be checked or unchecked. The peak window size shown in Figure 13 is not the default setting. It has been changed to 11 from the default value of 15. This ensures that the software can resolve alleles in the ladder that are 1 base apart for the larger loci such as D1S1656, D12S391 and D2S441. Promega Corporation · 2800 Woods Hollow Road · Madison, WI 53711-5399 USA · Toll Free in USA 800-356-9526 · 608-274-4330 · Fax 608-277-2516 · www.promega.com TMD041 Revised 10/23...
  • Page 39 Select Save to save the new analysis method. Select Done to exit the GeneMapper ID-X Manager. ® Promega Corporation · 2800 Woods Hollow Road · Madison, WI 53711-5399 USA · Toll Free in USA 800-356-9526 · 608-274-4330 · Fax 608-277-2516 TMD041 · Revised 10/23 www.promega.com...
  • Page 40 We recommend that you modify the values in these tabs to fit your laboratory’s data analysis protocols. Promega Corporation · 2800 Woods Hollow Road · Madison, WI 53711-5399 USA · Toll Free in USA 800-356-9526 · 608-274-4330 · Fax 608-277-2516 ·...

  • Page 41: Creating A Databasing Or Paternity Analysis Method With Genemapper ® Id-X Software

    Select the ‘Allele’ tab (Figure 15). Select the bins text file that was imported in Section 6.A. Promega Corporation · 2800 Woods Hollow Road · Madison, WI 53711-5399 USA · Toll Free in USA 800-356-9526 · 608-274-4330 · Fax 608-277-2516 TMD041 · Revised 10/23...
  • Page 42 Amelogenin Cutoff and Global Cutoff Value for Tri and Tetra repeats (Figure 15). In-house validation should be performed. Figure 15. The GeneMapper ID-X ‘Allele’ tab with settings for using a 20% peak filter. ® Promega Corporation · 2800 Woods Hollow Road · Madison, WI 53711-5399 USA · Toll Free in USA 800-356-9526 · 608-274-4330 · Fax 608-277-2516 · www.promega.com TMD041 Revised 10/23...
  • Page 43 In the ‘Size Standard’ column, select the size standard that was imported in Section 6.B or created in Section 6.C. Promega Corporation · 2800 Woods Hollow Road · Madison, WI 53711-5399 USA · Toll Free in USA 800-356-9526 · 608-274-4330 · Fax 608-277-2516 TMD041 · Revised 10/23...

  • Page 44: Importing Powerplex ® Esi Fast Panels And Bins Text Files Into Genemapper ® Id Software, Version 3.2

    Save the WEN_ILS_500_IDX.xml file to a known location on your computer. Promega Corporation · 2800 Woods Hollow Road · Madison, WI 53711-5399 USA · Toll Free in USA 800-356-9526 · 608-274-4330 · Fax 608-277-2516 ·...

  • Page 45: Importing The Wen Ils 500 Ess Size Standard Into Genemapper Id Software, Version 3.2

    Highlight the file, and then select Import. Select Done to save changes and exit the GeneMapper Manager. ® Promega Corporation · 2800 Woods Hollow Road · Madison, WI 53711-5399 USA · Toll Free in USA 800-356-9526 · 608-274-4330 · Fax 608-277-2516 TMD041 · Revised 10/23 www.promega.com...

  • Page 46: Creating A Size Standard With Genemapper ® Id Software, Version 3.2

    Select Basic or Advanced (Figure 16). The type of analysis method selected must match the type of analysis method created earlier. Select OK. Figure 16. The ‘Select Dye and Analysis Method’ window. Promega Corporation · 2800 Woods Hollow Road · Madison, WI 53711-5399 USA · Toll Free in USA 800-356-9526 · 608-274-4330 · Fax 608-277-2516 · www.promega.com...
  • Page 47 300, 325, 350, 375, 400, 425, 450, 475 and 500 bases). See Section 9.B, Figure 23. Select OK. Promega Corporation · 2800 Woods Hollow Road · Madison, WI 53711-5399 USA · Toll Free in USA 800-356-9526 · 608-274-4330 · Fax 608-277-2516 TMD041 · Revised 10/23...

  • Page 48: Creating A Casework Analysis Method With Genemapper ® Id Software, Version 3.2

    Enter a descriptive name for the analysis method, such as “PowerPlex ESI 17 Fast”. Select the ‘Allele’ tab (Figure 18). Promega Corporation · 2800 Woods Hollow Road · Madison, WI 53711-5399 USA · Toll Free in USA 800-356-9526 · 608-274-4330 · Fax 608-277-2516 ·...
  • Page 49 System. For an explanation of the proper usage and effects of these settings, refer to the Applied Biosystems user bulletin titled “Installation Procedures and New Features for GeneMapper ID Software 3.2”. Note: Some of these settings have been optimized and are different from the recommended settings in the user bulletin. You may need to optimize these settings. In-house validation should be performed. Promega Corporation · 2800 Woods Hollow Road · Madison, WI 53711-5399 USA · Toll Free in USA 800-356-9526 · 608-274-4330 · Fax 608-277-2516 TMD041 · Revised 10/23 www.promega.com...
  • Page 50 15. This ensures that the software can resolve alleles in the ladder that are 1 base apart for the larger loci, such as D1S1656, D12S391 and D2S441. Promega Corporation · 2800 Woods Hollow Road · Madison, WI 53711-5399 USA · Toll Free in USA 800-356-9526 · 608-274-4330 · Fax 608-277-2516 ·...
  • Page 51 Browse to the location of the run files. Highlight desired files, and then select Add to list followed by Add. Promega Corporation · 2800 Woods Hollow Road · Madison, WI 53711-5399 USA · Toll Free in USA 800-356-9526 · 608-274-4330 · Fax 608-277-2516 TMD041 ·...

  • Page 52: Creating A Databasing Or Paternity Analysis Method With Genemapper ® Id Software, Version 3.2

    For example, for a 20% filter enter “0.20” for the Amelogenin Cutoff and the Global Cutoff Value for Tri and Tetra repeats (Figure 20). Promega Corporation · 2800 Woods Hollow Road · Madison, WI 53711-5399 USA · Toll Free in USA 800-356-9526 · 608-274-4330 · Fax 608-277-2516 ·...
  • Page 53 You may need to optimize these settings. In-house validation should be performed. Figure 20. The GeneMapper ID ‘Allele’ tab with settings for using a 20% peak filter. ® Promega Corporation · 2800 Woods Hollow Road · Madison, WI 53711-5399 USA · Toll Free in USA 800-356-9526 · 608-274-4330 · Fax 608-277-2516 TMD041 · Revised 10/23 www.promega.com...
  • Page 54 In the ‘Size Standard’ column, select the size standard that was imported in Section 6.G or created in Section 6.H. Select Analyze (green arrow button) to start the data analysis. Promega Corporation · 2800 Woods Hollow Road · Madison, WI 53711-5399 USA · Toll Free in USA 800-356-9526 · 608-274-4330 · Fax 608-277-2516 · www.promega.com...

  • Page 55: Controls

    60bp to 500bp fragments of the WEN Internal Lane Standard 500 ESS. Promega Corporation · 2800 Woods Hollow Road · Madison, WI 53711-5399 USA · Toll Free in USA 800-356-9526 · 608-274-4330 · Fax 608-277-2516 TMD041 · Revised 10/23...
  • Page 56 ID software, version 3.2, and in the stutter text file for ® locus-specific filtering in GeneMapper ID-X software. ® Promega Corporation · 2800 Woods Hollow Road · Madison, WI 53711-5399 USA · Toll Free in USA 800-356-9526 · 608-274-4330 · Fax 608-277-2516 · www.promega.com TMD041 Revised 10/23...
  • Page 57 Number of bases below (n–) or above (n+) the true allele peak, respectively. Promega Corporation · 2800 Woods Hollow Road · Madison, WI 53711-5399 USA · Toll Free in USA 800-356-9526 · 608-274-4330 · Fax 608-277-2516 TMD041 · Revised 10/23...
  • Page 58 This artifact is not observed in no-amplification controls. Peak may be above or below analysis threshold depending on sensitivity of the capillary electrophoresis instrument. Promega Corporation · 2800 Woods Hollow Road · Madison, WI 53711-5399 USA · Toll Free in USA 800-356-9526 · 608-274-4330 · Fax 608-277-2516 ·...

  • Page 59: Troubleshooting

    2800M Control DNA is stored at +2°C to +10°C for at least 24 hours before use; do not refreeze. Vortex before use. Promega Corporation · 2800 Woods Hollow Road · Madison, WI 53711-5399 USA · Toll Free in USA 800-356-9526 · 608-274-4330 · Fax 608-277-2516 TMD041 · Revised 10/23...
  • Page 60 G5 spectral calibration is set for ® G5. See instructions for instrument preparation in Section 5. Promega Corporation · 2800 Woods Hollow Road · Madison, WI 53711-5399 USA · Toll Free in USA 800-356-9526 · 608-274-4330 · Fax 608-277-2516 · www.promega.com...
  • Page 61 Use a different injection of allelic ladder to determine sizes. Promega Corporation · 2800 Woods Hollow Road · Madison, WI 53711-5399 USA · Toll Free in USA 800-356-9526 · 608-274-4330 · Fax 608-277-2516 TMD041 · Revised 10/23...

  • Page 62: Amplification Of Extracted Dna

    25µl for extracted DNA. Decreasing the reaction volume may result in suboptimal performance. Promega Corporation · 2800 Woods Hollow Road · Madison, WI 53711-5399 USA · Toll Free in USA 800-356-9526 · 608-274-4330 · Fax 608-277-2516 · www.promega.com...
  • Page 63 25μl. Decreasing the reaction volume can result in suboptimal performance. Promega Corporation · 2800 Woods Hollow Road · Madison, WI 53711-5399 USA · Toll Free in USA 800-356-9526 · 608-274-4330 · Fax 608-277-2516 TMD041 · Revised 10/23...

  • Page 64: Direct Amplification Of Dna From Storage Card Punches

    Incubation for shorter time periods may result in incomplete inactivation of the PunchSolution™ Reagent. We have not tested longer incubation times. Promega Corporation · 2800 Woods Hollow Road · Madison, WI 53711-5399 USA · Toll Free in USA 800-356-9526 · 608-274-4330 · Fax 608-277-2516 · www.promega.com...
  • Page 65 Promega Corporation · 2800 Woods Hollow Road · Madison, WI 53711-5399 USA · Toll Free in USA 800-356-9526 · 608-274-4330 · Fax 608-277-2516 TMD041 · Revised 10/23...
  • Page 66 Review the amplification setup and confirm whether or not AmpSolution™ Reagent is required for your direct amplification reactions. Promega Corporation · 2800 Woods Hollow Road · Madison, WI 53711-5399 USA · Toll Free in USA 800-356-9526 · 608-274-4330 · Fax 608-277-2516 · www.promega.com...
  • Page 67 DNA present. Promega Corporation · 2800 Woods Hollow Road · Madison, WI 53711-5399 USA · Toll Free in USA 800-356-9526 · 608-274-4330 · Fax 608-277-2516 TMD041 · Revised 10/23 www.promega.com...

  • Page 68: Direct Amplification Of Dna From Swabs

    DNA was not accessible on nonlytic material. Pretreat swabs with SwabSolution™ Reagent to ensure that DNA is released from cellular proteins. Promega Corporation · 2800 Woods Hollow Road · Madison, WI 53711-5399 USA · Toll Free in USA 800-356-9526 · 608-274-4330 · Fax 608-277-2516 · www.promega.com...
  • Page 69 “shadow” peaks that differ in size by approximately the same distance as the single- stranded alleles. Promega Corporation · 2800 Woods Hollow Road · Madison, WI 53711-5399 USA · Toll Free in USA 800-356-9526 · 608-274-4330 · Fax 608-277-2516 TMD041 · Revised 10/23 www.promega.com...
  • Page 70 Pretreat swabs with SwabSolution™ Reagent to ensure that DNA is released from cellular proteins. Promega Corporation · 2800 Woods Hollow Road · Madison, WI 53711-5399 USA · Toll Free in USA 800-356-9526 · 608-274-4330 · Fax 608-277-2516 · www.promega.com...

  • Page 71: Genemapper ® Id-X Software

    Promega Corporation · 2800 Woods Hollow Road · Madison, WI 53711-5399 USA · Toll Free in USA 800-356-9526 · 608-274-4330 · Fax 608-277-2516 TMD041 · Revised 10/23...
  • Page 72 A low-quality allelic ladder was used during analysis. Ensure that only high-quality allelic ladders are used for analysis. Promega Corporation · 2800 Woods Hollow Road · Madison, WI 53711-5399 USA · Toll Free in USA 800-356-9526 · 608-274-4330 · Fax 608-277-2516 ·...

  • Page 73: Genemapper ® Id Software

    ‘Basic or Advanced’ as the analysis type. If they are different, an error is obtained. Promega Corporation · 2800 Woods Hollow Road · Madison, WI 53711-5399 USA · Toll Free in USA 800-356-9526 · 608-274-4330 · Fax 608-277-2516 TMD041 · Revised 10/23...
  • Page 74 Section 6.J or 6.I. Adjust the starting data point in the analysis method. Alternatively, use a full range for the analysis. Promega Corporation · 2800 Woods Hollow Road · Madison, WI 53711-5399 USA · Toll Free in USA 800-356-9526 · 608-274-4330 · Fax 608-277-2516 · www.promega.com...
  • Page 75 ‘Allele’ tab. Be sure to choose the appropriate bins text file, as shown in Figure 21. Promega Corporation · 2800 Woods Hollow Road · Madison, WI 53711-5399 USA · Toll Free in USA 800-356-9526 · 608-274-4330 · Fax 608-277-2516 TMD041 · Revised 10/23...
  • Page 76 The analysis type cannot be changed. If the method is not HID, it should be deleted and a new analysis method created. Promega Corporation · 2800 Woods Hollow Road · Madison, WI 53711-5399 USA · Toll Free in USA 800-356-9526 · 608-274-4330 · Fax 608-277-2516 ·...

  • Page 77: References

    PCR Protocols: A Guide to Methods and Applications (1990) Innis, M.A. et al. eds., Academic Press, San Diego, Butler, J.M. (2005) Forensic DNA Typing, 2nd ed. Elsevier Academic Press, London. Promega Corporation · 2800 Woods Hollow Road · Madison, WI 53711-5399 USA · Toll Free in USA 800-356-9526 · 608-274-4330 · Fax 608-277-2516 TMD041 · Revised 10/23...
  • Page 78 Int. J. Legal Med. 110, 175–6. Gill, P. et al. (1997) Considerations from the European DNA Profiling Group (EDNAP) concerning STR nomenclature. Forensic Sci. Int. 87, 185–92. Promega Corporation · 2800 Woods Hollow Road · Madison, WI 53711-5399 USA · Toll Free in USA 800-356-9526 · 608-274-4330 · Fax 608-277-2516 · www.promega.com...

  • Page 79: Appendix

    60°C (20) to the amplification protocols to provide conditions for essentially complete terminal nucleotide addition when recommended amounts of template DNA are used. Promega Corporation · 2800 Woods Hollow Road · Madison, WI 53711-5399 USA · Toll Free in USA 800-356-9526 · 608-274-4330 · Fax 608-277-2516 TMD041 · Revised 10/23...
  • Page 80 Amelogenin is not an STR. NA = Not applicable. Promega Corporation · 2800 Woods Hollow Road · Madison, WI 53711-5399 USA · Toll Free in USA 800-356-9526 · 608-274-4330 · Fax 608-277-2516 · www.promega.com...
  • Page 81 Technology (NIST) web site at: https://strbase.nist.gov/var_tab.htm Amelogenin is not an STR. Promega Corporation · 2800 Woods Hollow Road · Madison, WI 53711-5399 USA · Toll Free in USA 800-356-9526 · 608-274-4330 · Fax 608-277-2516 TMD041 · Revised 10/23 www.promega.com...
  • Page 82 16, 19 TH01 6, 9.3 SE33 15, 16 20, 23 D8S1179 14, 15 Promega Corporation · 2800 Woods Hollow Road · Madison, WI 53711-5399 USA · Toll Free in USA 800-356-9526 · 608-274-4330 · Fax 608-277-2516 · www.promega.com TMD041 Revised 10/23...

  • Page 83: The Wen Internal Lane Standard 500 Ess

    1.2mm Harris Micro-Punch or equivalent manual punch and cutting mat or automated punch system Promega Corporation · 2800 Woods Hollow Road · Madison, WI 53711-5399 USA · Toll Free in USA 800-356-9526 · 608-274-4330 · Fax 608-277-2516 TMD041 · Revised 10/23...
  • Page 84 PunchSolution™ Reagent to the well before adding the punch during pretreatment may help alleviate static problems. Promega Corporation · 2800 Woods Hollow Road · Madison, WI 53711-5399 USA · Toll Free in USA 800-356-9526 · 608-274-4330 · Fax 608-277-2516 ·...
  • Page 85 ESI 17 Fast 10X Primer Pair Mix and 5X AmpSolution™ Reagent. For lytic storage card punches, the template DNA will be added at Step 7. Promega Corporation · 2800 Woods Hollow Road · Madison, WI 53711-5399 USA · Toll Free in USA 800-356-9526 · 608-274-4330 · Fax 608-277-2516 TMD041 · Revised 10/23...
  • Page 86 Thermal Cycling Amplification and detection instrumentation may vary. You will need to optimize protocols, including cycle number, injection conditions and loading volume for your laboratory instrumentation. Testing at Promega shows that 25 cycles works well for a variety of storage card sample types. Cycle number will need to be optimized in each laboratory for each sample type. Note: It may be possible to use thermal cyclers other than those listed in this technical manual. Use of thermal cyclers not listed here may require optimization of cycling conditions and validation in your laboratory. Use of thermal cyclers with an aluminum block is not recommended with the PowerPlex ESI 17 Fast System. ® Promega Corporation · 2800 Woods Hollow Road · Madison, WI 53711-5399 USA · Toll Free in USA 800-356-9526 · 608-274-4330 · Fax 608-277-2516 · www.promega.com TMD041 Revised 10/23...
  • Page 87 Following amplification, use your laboratory’s validated separation and detection protocols to determine the optimal cycle number for the sample type. Promega Corporation · 2800 Woods Hollow Road · Madison, WI 53711-5399 USA · Toll Free in USA 800-356-9526 · 608-274-4330 · Fax 608-277-2516 TMD041 · Revised 10/23...

  • Page 88: Direct Amplification Of Dna From Swabs In A 12.5Μl Reaction Volume

    Use a clean plate for reaction assembly, and label it appropriately. Alternatively, determine the number of clean, 0.2ml reaction tubes required, and label them appropriately. Promega Corporation · 2800 Woods Hollow Road · Madison, WI 53711-5399 USA · Toll Free in USA 800-356-9526 · 608-274-4330 · Fax 608-277-2516 ·...
  • Page 89 Note: Optimization of the amount of 2800M Control DNA may be required depending on thermal cycling conditions and laboratory preferences. For the negative amplification control, pipet 2µl of Water, Amplification Grade, or TE buffer instead of swab –4 extract into a reaction well containing PCR amplification mix. Note: Additional negative controls can be included. Assemble a reaction containing the swab extract prepared from a blank swab, or assemble a reaction where the SwabSolution™ Reagent is processed as a blank without a swab. 10. Seal or cap the plate, or close the tubes. Optional: Briefly centrifuge the plate to bring contents to the bottom of the wells and remove any air bubbles. Promega Corporation · 2800 Woods Hollow Road · Madison, WI 53711-5399 USA · Toll Free in USA 800-356-9526 · 608-274-4330 · Fax 608-277-2516 TMD041 · Revised 10/23 www.promega.com...
  • Page 90 Thermal Cycling Protocol 96°C for 1 minute, then: 96°C for 5 seconds 60°C for 35 seconds 72°C for 5 seconds for 25 cycles, then: 60°C for 2 minutes 4°C soak After completion of the thermal cycling protocol, proceed with fragment analysis or store amplified samples at –20°C protected from light. Note: Long-term storage of amplified samples at 4°C or higher may produce artifacts. Promega Corporation · 2800 Woods Hollow Road · Madison, WI 53711-5399 USA · Toll Free in USA 800-356-9526 · 608-274-4330 · Fax 608-277-2516 · www.promega.com TMD041 Revised 10/23...

  • Page 91: Composition Of Buffers And Solutions

    Dissolve Tris base and EDTA in 900ml of deionized water. Adjust to pH 8.0 with HCl. Add glycogen. Bring the final volume to 1 liter with deionized water. Promega Corporation · 2800 Woods Hollow Road · Madison, WI 53711-5399 USA · Toll Free in USA 800-356-9526 · 608-274-4330 · Fax 608-277-2516 TMD041 · Revised 10/23...

  • Page 92: Related Products

    © 2009–2023 Promega Corporation. All Rights Reserved. PowerPlex is a registered trademark of Promega Corporation. AmpSolution, PunchSolution and SwabSolution are trademarks of Promega Corporation. Applied Biosystems is a registered trademark of Applied Biosystems LLC. Bode Buccal DNA Collector is a trademark of the Bode Technology Group, Inc. EasiCollect and OmniSwab are trademarks of Whatman. FTA is a registered trademark of Flinders Technologies, Pty, Ltd., and is licensed to Whatman. GeneAmp, GeneMapper, MicroAmp, POP-4, ProFlex and Veriti are registered trademarks and GeneScan, Hi-Di and POP-6 are trademarks of Thermo Fisher Scientific. Vacutainer is a registered trademark of Becton, Dickinson and Company. Products may be covered by pending or issued patents or may have certain limitations. Please visit our Web site for more information. All prices and specifications are subject to change without prior notice. Product claims are subject to change. Please contact Promega Technical Services or access the Promega online catalog for the most up-to-date information on Promega products. Promega Corporation · 2800 Woods Hollow Road · Madison, WI 53711-5399 USA · Toll Free in USA 800-356-9526 · 608-274-4330 · Fax 608-277-2516 · www.promega.com TMD041 Revised 10/23...

Table of Contents