Rapid Solutions for Processing Swabs & Punches PunchSolution and SwabSolution Kits 5X AmpSolution Reagent

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1 Rapid Solutions for Processing Swabs & Punches PunchSolution and SwabSolution Kits 5X AmpSolution Reagent May 2012

2 Faster Sample Processing Database, Paternity and Reference Samples Rapid methods available from unwashed FTA punches Direct amplification STR kits Hours can be saved in analysis of each plate Benefits great for FTA users, but other samples are common Swabs? S&S903 card punches, etc.? Non-direct STR kits?

3 Buccal Swabs and Non-FTA Cards Direct Amplification Problem Buccal Swabs Too big to punch using automated punchers No lysis or protein denaturation (DNA accessibility issue) DNA may not be evenly distributed over surface of swab Extraction and purification time consuming and expensive Non-FTA Punches Can use manual and automated punchers, but No lysis or protein denaturation (DNA accessibility issue)

4 Buccal Swabs and Non-FTA Cards Rapid Processing Solutions SwabSolution Kit Quick, simple extraction for swabs PunchSolution Kit Quick, simple pre-processing step for punches from non-fta cards Lyse cells and deproteinate to make DNA accessible Produce robust PowerPlex signal, even from old samples

5 SwabSolution and PunchSolution Kits For use with PowerPlex Systems SwabSolution Kit (DC8271) 100 preps Contains: SwabSolution Reagent, 100mL 5X AmpSolution Reagent, 500µL* PunchSolution Kit (DC9271) 100 preps Contains: PunchSolution Reagent, 1mL 5X AmpSolution Reagent, 500µL* * 5X AmpSolution Reagent Added to PowerPlex reaction mix Used in PowerPlex 16, 16 HS, ESX, ESI Systems Not used with PowerPlex 18D, 21, Y23 Systems Available separately (DM1231)

6 Rapid Processing of Non-FTA Punches PunchSolution Kit for PowerPlex Systems

7 PunchSolution Kit Non-FTA punch workflow (S&S903, Bode Buccal DNA Collector) Rapid, same-plate processing of non-fta punches For PowerPlex reactions other than PowerPlex 18D, 21 and Y23 Systems, add AmpSolution Reagent to reaction mix

8 PunchSolution Kit Blood on S&S 903, PowerPlex 18D Amplification 1.2mm punch pretreated with PunchSolution Kit; PowerPlex 18D System (27 cycles)

9 PunchSolution vs. Bode PunchPrep vs. Untreated PunchSolution Produces Robust Peaks Untreated Untreated BPP BPP PS PS Untreated Untreated BPP BPP PS PS BPP: Bode PunchPrep treated PS: Promega PunchSolution treated 1.2mm punch from Bode Buccal DNA Collector; PowerPlex 18D System (27 cycles)

10 Rapid Processing of Swabs SwabSolution Kit for PowerPlex Systems

11 SwabSolution Kit Buccal Swab Workflow Simple, rapid process for all swabs * For plate format, use 2.2mL deep well plate (V6781) ** Use new heat transfer block (A2661) For reactions other than PowerPlex 18D and PowerPlex 21, add AmpSolution Reagent

12 SwabSolution Kit OmniSwab Extract PowerPlex 16 PowerPlex 16 HS 2µL of SwabSolution extract; PowerPlex 16 and 16 HS System with AmpSolution Reagent (28 cycles)

13 Data Quality SwabSolution vs. DNA IQ Extracted DNA SwabSolution SwabSolution DNA IQ DNA IQ SwabSolution SwabSolution DNA IQ DNA IQ OmniSwab ; PowerPlex 18D System Equivalent balance obtained with SwabSolution and DNA IQ extracts Strong PowerPlex profiles from fresh and old swabs

14 SwabSolution Kit DNA can be stored for future use Stable at 4 C (current data out to 6 months) Quantification is possible, if desired PicoGreen or QuantiFluor (DNA is double-stranded) Plexor HY (with AmpSolution Reagent in reaction mix)

15 Quantitation with SwabSolution Extracts PicoGreen QuantiFluor Plexor HY Swab #1 6.8ng/uL 8.2ng/uL 7.2ng/uL Swab #2 8.8ng/uL 10.7ng/uL 10.2ng/uL Swab #3 5.9ng/uL 7.7ng/uL 8.4ng/uL Swab #4 4.9ng/uL 6.0ng/uL 9.8ng/uL Compatible with Quant-iT PicoGreen dsdna Reagent, QuantiFluor dsdna System and Plexor HY System Allows normalization, if desired (not required) Data show ample yield for PowerPlex analysis from buccal samples

16 Optional Normalization Based on PicoGreen Data Average Peak Height Average Peak Height Non-Normalized Normalized: 5ng/25uL Swab#1 Swab#2 Swab#3 Swab#4 PowerPlex 18D System

17 Direct Amplification from FTA Punches New application for traditional PowerPlex Systems

18 Direct Amplification from FTA Punches Additional Application for AmpSolution Reagent Not all PowerPlex Systems were designed for direct amplification from FTA punches 5X AmpSolution Reagent (DM1231) allows direct amp with FTA for some traditional PowerPlex Systems PowerPlex ESX/ESI, 16 HS, CS7 Systems AmpSolution is simply added to the master mix Provides a new option for current PowerPlex users

19 Direct Amplification from Blood FTA Punches PowerPlex 16 HS System +AS +AS -AS -AS +AS 1.2mm punch from blood FTA card; PowerPlex 16 HS System (27 cycles) +AS: AmpSolution in reaction mix AS: No AmpSolution -AS Good results with PowerPlex 16 HS Testing of other PowerPlex Systems underway

20 Direct Amplification from Buccal FTA Punches PowerPlex 16 HS System +AS +AS -AS -AS +AS 2 x 1.2mm punch from buccal FTA card; PowerPlex 16 HS System (27 cycles) -AS Provides faster FTA processing for current PowerPlex users +AS: AmpSolution in reaction mix AS: No AmpSolution

21 Direct Amplification from FTA Punches PowerPlex ESX and ESI Systems PowerPlex ESI 17 System Buccal PowerPlex ESX 17 System Buccal Blood Blood

22 Simple Workflows for Database & Paternity Testing Rapid sample processing for PowerPlex Systems PunchSolution Kit for non-fta punches SwabSolution Kit for buccal swabs 5X AmpSolution Reagent for above Kits or direct amplification from FTA with traditional PowerPlex Systems No matter your sample type, do it faster and without compromise in profile quality

23 Acknowledgements R&D Bob McLaren Jay Patel Katie Oostdik Doug Storts Manufacturing Katie Hebble Jennifer Setlak Marketing Ben Krenke Sales & Support Kim Huston Rita Weispfenning Project Management Susan Frackman Scientific Communications Kelly Grooms Terri Sundquist Quality Assurance Susan Wigdal

24 Questions Contact your local Promega representative or for technical support. Visit to learn more.