Hot-start DNA polymerases are commonly used in PCR for genotyping, sequencing, molecular diagnostics, and high-throughput applications. In this presentation, PCR performance of Invitrogen™ Platinum II Taq Hot-Start DNA Polymerase and Invitrogen™ AccuPrime Taq DNA Polymerase is compared in the following areas: • PCR run time for targets of different lengths • Amplification of AT-rich and GC-rich sequences • Tolerance to PCR inhibitors • Sensitivity in target detection • Universal protocol for PCR targets of different lengths • Multiplex PCR of 15 targets • Product format for direct gel loading Request a sample of Platinum II Taq enzyme at http://bit.ly/2M4U9cw Find other PCR enzymes at http://bit.ly/2JIPrzj Learn more about PCR at http://bit.ly/2y2aSVo #PCR #PCREducation #Invitrogen #InvitrogenSchoolofMolBio

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Powerful hot-start DNA polymerase:
PCR performance comparison of Platinum II Taq and AccuPrime Taq enzymes
For Research Use Only. Not for use in diagnostic procedures.

2. 2
PCR results are obtained ≥2 times faster with the Platinum II Taq enzyme than with the AccuPrime Taq enzyme.
Comparison of PCR Run Time
0
20
40
60
80
100
120
140
160
180
200 PlatinumIITaq
AccuPrimeTaq
PlatinumIITaq
AccuPrimeTaq
PlatinumIITaq
AccuPrimeTaq
500 bp fragment 2 kb fragment 4 kb fragment
PCRruntime(min)
Ramping time Cycling time
Short PCR run time from fast cycling
500 bp, 2 kb, and 4 kb fragments were amplified from
human genomic DNA (gDNA) using Invitrogen™
Platinum™ II Taq Hot-Start DNA Polymerase or
Invitrogen™ AccuPrime™ Taq DNA Polymerase. PCR
was set up in 50 μL reactions for 35 cycles. Cycling
times for both polymerases are shown in dark blue,
while ramping times on the Applied Biosystems™
ProFlex™ PCR System (6°C/sec peak block ramp
rate) are shown in gray.
PlatinumIITaq
AccuPrimeTaq
PlatinumIITaq
AccuPrimeTaq
PlatinumIITaq
AccuPrimeTaq

3. 3
Amplification of AT-Rich and GC-Rich Targets
Targets with higher GC content (>67%) were amplified successfully
with Platinum II Taq DNA Polymerase supplemented with Platinum GC Enhancer.
PlatinumII
Taqenzyme
AccuPrime
Taqenzyme
GC (%)
29 30 35 41 57 60 66 67 69 70 71 73 74 76 77
Robust amplification of high-AT and
high-GC targets
DNA fragments of varying GC content (indicated
above the corresponding lanes) were amplified using
Platinum II Taq Hot-Start DNA Polymerase or
AccuPrime Taq DNA Polymerase. 100 ng of human
gDNA were used as templates in 50 µL PCR reactions.
Invitrogen™ Platinum™ GC Enhancer was
supplemented for targets with >65% GC. Ladder:
Thermo Scientific™ ZipRuler™ Express DNA Ladder 2.

4. 4
Humic
acid
Hemin Xylan Humic
acid
Hemin Xylan
Tolerance to Common PCR Inhibitors
High tolerance to
inhibitors
A 1 kb fragment of human gDNA
was amplified using Platinum II Taq
Hot-Start DNA Polymerase or AccuPrime
Taq DNA Polymerase. The reactions were
spiked in with humic acid (found in soil),
hemin (found in blood), or xylan (found in
plants) at the final concentrations
indicated. Ladder: Thermo Scientific™
ZipRuler™ Express DNA Ladder 1.
Higher tolerance to common PCR inhibitors was observed with Platinum II Taq DNA polymerase.
Platinum II Taq enzyme AccuPrime Taq enzyme
0 1 2
μg/mL
0 10 20
μM
0 0.3 0.6
mg/mL
0 1 2
μg/mL
0 10 20
μM
0 0.3 0.6
mg/mL

5. 5
Sensitivity in Target Detection
Equal or better sensitivity was observed with Platinum II Taq enzyme compared to
AccuPrime Taq enzyme, depending on the target length.
• Similar sensitivity with <1 kb fragments
• Better sensitivity with 1–2 kb fragments
0 5 10 20 40 80 160 320
(copies of human gDNA)
0 5 10 20 40 80 160 320
(copies of human gDNA)
0 5 10 20 40 80 160 320
(copies of human gDNA)
0 5 10 20 40 80 160 320
(copies of human gDNA)
Platinum II Taq enzyme AccuPrime Taq enzyme
High sensitivity in detection of
low DNA input
595 bp and 1,988 bp fragments were amplified from
varying amounts of human gDNA using Platinum II Taq
Hot-Start DNA Polymerase or AccuPrime Taq DNA
Polymerase. Estimated copy numbers of input DNA in
50 μL PCR reactions are indicated. 0.0016 ng of human
gDNA equals about 5 copies. Ladder: ZipRuler Express
DNA Ladder 2.
595 bp
1,988 bp

6. 6
Platinum II Taq enzyme AccuPrime Taq enzyme
Using Same PCR Protocol for Different Targets
Different PCR assays can be cycled together using the same universal protocol
using Platinum II Taq DNA polymerase, enabling significant time savings.
132bp
251bp
735bp
1kb
3.9kb
132bp
251bp
735bp
1kb
3.9kb
Universal protocol for
different PCR targets
Fragments of 132 bp, 251 bp, 735 bp, 1 kb, and 3.9 kb
were amplified individually from 50 ng human gDNA
using Platinum II Taq Hot-Start DNA Polymerase or
AccuPrime Taq DNA Polymerase. PCR was set up in
50 µL reactions for 35 cycles. The annealing
temperature, extension time, and other cycling
parameters of the 3.9 kb fragment were followed for all
fragments amplified. Ladder: ZipRuler Express DNA
Ladder Express 1.
60°C, 15 sec
68°C, 1 min
54°C, 30 sec
68°C, 4 min
Annealing and extension
3.9 kb cycling for all targets 3.9 kb cycling for all targets

7. 7
Running Multiplex PCR
Multiple targets, including longer fragments, can be amplified in the same reaction
using Platinum II Taq enzyme, contrary to AccuPrime Taq enzyme.
Enabling multiplex PCR
15 targets of increasing sizes (99 bp, 131 bp, 149 bp, 199 bp,
251 bp, 297 bp, 345 bp, 400 bp, 516 bp, 613 bp, 735 bp, 908 bp,
1,005 bp, 1,190 bp, and 1,606 bp) were simultaneously amplified
from 200 ng and 1,000 ng of human gDNA using Platinum II Taq
Hot-Start DNA Polymerase or AccuPrime Taq DNA Polymerase.
PCR was set up in 50 μL reactions. Ladder: ZipRuler Express DNA
Ladder Express 2.
200ng
200ng
1,000ng
1,606 bp
1,190 bp
1,005 bp
908 bp
735 bp
613 bp
516 bp
400 bp
345 bp
297 bp
251 bp
199 bp
149 bp
131 bp
99 bp
Platinum II
Taq enzyme
AccuPrime
Taq enzyme
1,000ng

8. 8
The green buffer options are available as stand-alone and master mix product formats
for the Platinum II Taq enzyme, but not for the AccuPrime Taq enzyme.
Availability of Direct Gel-Loading Formats
Green buffer formats
The green buffer options allow for direct gel loading to track DNA
migration with two dyes during electrophoresis. The green buffer
format is also available for master mixes. Green buffer product
formats with direct gel loading help save time, prevent pipetting
errors, and reduce waste in PCR.

9. 9
Summary Comparison of Platinum II Taq and AccuPrime Taq DNA Polymerases
Platinum II Taq Hot-Start DNA Polymerase
AccuPrime Taq
DNA Polymerase
Hot-start modification Antibody-mediated Antibody-mediated
DNA synthesis rate 15 sec/kb 1 min/kb
Universal annealing protocol Yes No
Sensitivity +++ ++
Specificity +++ +++
GC-rich amplification +++ +
Inhibitor tolerance ++++ ++
Amplicon length Up to 5 kb Up to 4 kb
Fidelity (vs. Taq enzyme) 1x 1x
Master mix format Yes Yes
Formats for direct gel loading Yes No
Request a sample of Platinum II Taq enzyme at thermofisher.com/platinumiitaq*
* Terms and conditions apply.

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For Research Use Only. Not for use in diagnostic procedures. © 2019 Thermo Fisher Scientific Inc. All rights reserved.
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