WellAware™ Forensic Package

1. WellAware™ Forensic Package for use with…

2. Settings MenuUsed to allow customization for any lab.

3. WellAware™ Forensic Package Flow Diagram ABI 310/3100 Sequencer WellAware™ Complete WellAware™ Compact ABI 7000 RT-PCR

4. WellAware™ Complete WellAware™ Forensic Package WellAware™ Forensic Package ABI 7000 RT-PCR 1 WellAware™ Forensic Package How it Works 1. Samples that have been extracted and are ready for quantification with an AB 7000 Real Time PCR instrument, are entered into the plate form and then printed. 2. The next step is to export a file for the quantification setup in WellAware format. This file is opened in WellAware which then guides the scientist to load the extracted samples into the AB 7000 Real Time PCR plate. WellAware voice and light cues direct the addition of master mix to the wells. 3. In the Forensic Interface, the scientist exports plate format information for running the AB 7000. 4. Once the AB 7000 real time PCR instrument has completed the quantification of samples, the results from the AB 7000 Prism Instrument software are imported back into the Forensic Interface. Plate Form for ABI 7000 2 3 4

5. ABI 310/3100 Sequencer WellAware™ Forensic Package WellAware™ Compact Processing Screen Norm & 310/3100 data 5. The DNA processing screen allows the scientist to add the samples that have now been quantitated to an AB 310 or AB 3100 plate for electrophoresis and detection. This set up is flexible, and the scientist can run a sample multiple times, and can even override the dilution amount if they choose. Additional sample descriptions, for example case number, can be added under the sample header. 6. Once the scientist has the samples in the order they have specified, they can just click on either the AB 310 or AB 3100 tab, depending upon which platform they would like to set up. After printing the sample set-up sheet, a WellAware file can be created by clicking on the Export Data tab and designating whether to pipette by rows or columns. 7. The exported file is opened in the WellAware software and the scientist can now set up their dilution plate for amplification. WellAware calls out how much dilutant (TE or H2O) to add to each well of the dilution plate. It will then ask for the barcoded extracts to be scanned. Once scanned, WellAware calls out the volume of DNA to add to each well of the dilution plate, and lights up the destination well. Once the sample has been pipetted the footswitch is used to proceed to the next sample. 8. The final step in using the Forensic Interface is in the exporting of the AB 310 or AB 3100 file. The information put into the processing screen for the dilution, step 6, is used to generate the file for the AB 310 or AB 3100 Genetic Analyzers. WellAware™ Forensic Package How it Works 5 8 6 7

6. WellAware™ Forensic Package Main Screen Step 5 Step 3 Step 2 Step 1 Step 7 Step 6 Step 8 Step 8 Step 4

7. Quant Worksheet Step 1

8. WellAware™ Quant RT-PCR Pipetting Data (Page 1) Step 2

9. WellAware™ Quant RT-PCR Pipetting Data (Page 2)

10. WellAware™ Quant Pipetting a Step 3 b c

11. ABI 7000 real-time PCR Export Data Step 4

12. ABI 7000 RT-PCR system Normalization Data Step 6 Calculated normalization values Raw data from ABI 7000

13. Normalization Process Step 5 Step 7 Step 6 Pull down menu

14. WellAware™ Normalization Pipetting Data Step 7

15. ABI 310/3100 (Genetic Analyzer) Worksheet Step 7 Step 8

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