Download VersaTaq™ Direct PCR Polymerase

VersaTaq™ Direct PCR Polymerase
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VersaTaq Direct PCR Polymerase is a mutant Taq DNA polymerase engineered for direct amplification from complex inhibitory
templates.
n Eliminate extraction and purification steps
n Amplify limited quantities of DNA
n Increased sensitivity
n Amplifies from a wide variety of sample types
n Expedited experiments and data output
In contrast to standard Taq DNA polymerase, VersaTaq Direct
PCR Polymerase can amplify DNA directly from samples, which
eliminates costly extraction and purification steps, saving time
and enabling amplification from limited quantities of DNA. This
versatile polymerase remains functional in the presence of high
concentrations of inhibitors where other commercial enzymes
fail. Due to the wide variety of sample types supported by
VersaTaq Direct PCR Polymerase, optimization may be required.
VersaTaq Direct PCR Polymerase has 5’-3’ exonuclease activity,
making it suitable for hydrolysis probe-based detection chemistries.
Resistance to potent PCR inhibitors in a wide range of
end-point PCR applications:
– Whole blood (up to 40%)
– Feces
– Blood spots on sample – Urine
collection cards
– Animal tissue
– Buccal swabs
– Plant tissue
– Soil
– Crude extracts
Tissue:
Tissue, such as that isolated from genetically modified mouse
models, is extensively used in biomedical research. PCR from tissue samples typically requires a DNA extraction step due to PCR
inhibitors(1-2). VersaTaq Direct PCR Polymerase overcomes PCR
inhibitors commonly found in tissue, enabling direct amplification
of a wide range of DNA targets (Fig. 2).
Buccal swabs:
Buccal swabs for DNA collection can be used in a wide range of
applications such as genotyping and epigenetic studies. Buccal
swabs were rapidly eluted and added directly to PCR reactions
with VersaTaq Direct PCR Polymerase for amplification in a wide
range of targets (Fig. 3).
Blood spots:
VersaTaq Direct PCR Polymerase can amplify directly from blood
spotted on Whatman™ FTA™ elute cards (Fig. 4). Using VersaTaq
Direct PCR Polymerase decreases processing time significantly
because there is no need to elute DNA from the cards prior to
performing PCR.
Soil:
Environmental and forensic applications that require samples collected from crude soil for DNA amplification may present problems in PCR reactions. Trace amounts of humic acid commonly
found in aquatic soil and sediment environments have been
shown to inhibit PCR(3). VersaTaq Direct PCR Polymerase overcomes potent inhibitors, enabling direct amplification from a wide
range of targets, including those with high GC content (Fig. 5).
Whole blood:
VersaTaq Direct PCR Polymerase is able to amplify DNA in whole
blood (up to 40%) with various anticoagulants from a wide
range of targets including high GC (Fig. 1).
Source:
Recombinant
Blood (%)
Anticoagulant
M
Fig. 1. Amplification of targets directly from human whole blood.
E
10
H C
E
40
H C
E
10
H C
E
40
H C
E
10
H C
2000 bp -
E: Na-EDTA
H: Na-Heparin
C: Na-Citrate
500 bp -
M
1
2
3
4
5
6
7
8
9
10 11 12
13
14 15
Targets were amplified from different concentrations of human whole blood in the presence of varying types of anticoagulants. Lanes 1-6: NUMB 455 bp; Lanes
7-12: NUMB 1.8 kb; Lanes 13-15: HRES1 488 bp -corresponding band is an 81% GC amplicon. 1 M betaine added to reactions with high GC targets.
Concentration:
1.0 ± 0.2 µg/µl, determined by OD280
References:
1. Eckhart, L., Bach, J., Ban, J., and Tschachler, E. (2000) Biochem. Biophys. Res. Commun.
271, 726–730.
2. Kim, S., Labbe, R. G., and Ryu, S. (2000) Appl. Environ. Microbiol. 66, 1213–1215.
3. Tsai, Y.-L., and Olson, B. H. (1992) Appl. Environ. Microbiol. 58, 754-757.
Storage:
Shipped on dry ice. Store at -20°C.
71002
50 reactions
Anticoagulant
200 reactions
2000 bp -
FTA Cards
E
H
C
gDNA
Pack size
M
Product code
gDNA
Fig. 4. Amplification of targets directly from blood spots
VersaTaq Direct PCR Polymerase
FTA Cards
E
H
C
500 reactions
Fig. 2. Amplification of targets directly from mouse tissue
M
1
2
3
4
5
6
7
8
Amplification of targets from human whole blood spotted on Whatman
FTA elute cards by VersaTaq Direct PCR Polymerase. NUMB gene was
amplified from FTA cards spotted with human whole blood in the presence of varying types of anticoagulant and from genomic DNA control.
2 mm punches from FTA cards were placed directly into PCR reactions.
Lanes 1-4: NUMB 455 bp; Lanes 5-8: NUMB 967 bp.
1 mm
M
Mouse Tail (length)
E: Na-EDTA
H: Na-Heparin
C: Na-Citrate
500 bp -
2 x 500 reactions
2000 bp-
500 bp-
1
Fig. 5. Amplification of targets in presence of humic acid
2
Amplification of targets directly from mouse tail by VersaTaq Direct PCR
Polymerase. Targets were amplified from 1 mm mouse tail clip added
directly into PCR reaction. Lane 1: NUMB 483 bp; Lane 2: NUMB 1.8 kb.
Amplicon GC %
Humic acid (ng/µl)
M
M
48%
0.4 4
37%
0.4 4
81%
75%
0.4 1.6 0.4 1.6
M
1
3
5
2000 bp-
500 bp-
Amplicon length (kb)
M
Fig. 3. Amplification of targets directly from buccal swabs
0.2 0.3 0.4 0.5 0.6
1
2
2000 bp -
2
4
6
7
8
Amplification of diverse targets in presence of humic acid by VersaTaq
Direct PCR Polymerase. Targets were amplified from 5 ng of genomic DNA
in the presence of varying concentrations of humic acid. Lanes 1-2: NUMB
455 bp; Lanes 3-4: NUMB 1.8 kb; Lanes 5-6: HRES1 488 bp; Lanes 7-8:
SIM2 1.2 kb. 1 M betaine added to reactions with high GC content.
500 bp -
M
1
2
3
4
5
6
7
Amplification of targets directly from buccal swab eluent by VersaTaq
Direct PCR Polymerase. Targets were amplified from buccal swabs eluted
in 0.1% SDS and added directly into PCR reaction. Lane 1-7: NUMB 196
bp, 304 bp, 404 bp, 493 bp, 609 bp, 967 bp, 2 kb.
For Research Use Only. Not for use in diagnostic procedures.
Affymetrix and USB are registered trademarks of Affymetrix, Inc. VersaTaq is a trademark of Affymetrix. All other trademarks are the property of their respective owners.
Isogen Life Science B.V.
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3454 PW De Meern
The Netherlands
Tel: +31 (0)30 688 0771
Fax:+31 (0)30 688 8009
E-mail: [email protected]
© 2015 Affymetrix, Inc. All rights reserved.
P/N USB05287 Rev. 1
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Tel: +32 (0)9 351 48 79
Fax:+32 (0)9 277 93 03
Spain
Tel: +34 936 644 894
Fax: +34 936 455 089
www.isogen-lifescience.com
Rev 08/10/15