Rapid Solutions for Processing Swabs & Punches
PunchSolution™ and SwabSolution™ Kits
5X AmpSolution™ Reagent
May 2012
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?
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)
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
SwabSolution™ and PunchSolution™ Kits
For use with PowerPlex® Systems
SwabSolution™ Kit (DC8271)
PunchSolution™ Kit (DC9271)
Contains:
Contains:
• SwabSolution™ Reagent, 100mL
• 5X AmpSolution™ Reagent, 500µL*
• PunchSolution™ Reagent, 1mL
• 5X AmpSolution™ Reagent, 500µL*
100 preps
100 preps
* 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)
Rapid Processing of Non-FTA® Punches
PunchSolution™ Kit for PowerPlex® Systems
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
PunchSolution™ Kit
Blood on S&S 903, PowerPlex® 18D Amplification
1.2mm punch pretreated with PunchSolution™ Kit; PowerPlex® 18D System (27 cycles)
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)
Rapid Processing of Swabs
SwabSolution™ Kit for PowerPlex® Systems
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
SwabSolution™ Kit
OmniSwab™ Extract
PowerPlex® 16
PowerPlex®16 HS
2µL of SwabSolution extract;
PowerPlex® 16 and 16 HS System with
AmpSolution™ Reagent (28 cycles)
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
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)
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
Average Peak Height
Average Peak Height
Optional Normalization Based on PicoGreen® Data
8000
7000
6000
5000
4000
3000
2000
1000
0
5000
4000
3000
2000
1000
0
PowerPlex® 18D System
Non-Normalized
Swab#1
Swab#2
Swab#3
Swab#4
Normalized: 5ng/25uL
Direct Amplification from FTA® Punches
New application for traditional PowerPlex® Systems
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
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)
Good results with PowerPlex® 16 HS
-AS
+AS: AmpSolution™ in reaction mix
–AS: No AmpSolution™
Testing of other PowerPlex® Systems
underway
Direct Amplification from Buccal FTA® Punches
PowerPlex® 16 HS System
+AS
+AS
-AS
-AS
+AS
-AS
+AS: AmpSolution™ in reaction mix
–AS: No AmpSolution™
2 x 1.2mm punch from buccal FTA® card;
PowerPlex® 16 HS System (27 cycles)
Provides faster FTA® processing for
current PowerPlex® users
Direct Amplification from FTA® Punches
PowerPlex® ESX and ESI Systems
PowerPlex® ESI 17 System
PowerPlex® ESX 17 System
Buccal
Buccal
Blood
Blood
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
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
Questions
Contact your local Promega representative or
email [email protected] for technical support.
Visit www.promega.com/directamp to learn more.