JumpStart REDTaq DNA Polymerase

Catalog Number D8187

Storage Temperature –20 °C

TECHNICAL BULLETIN

Product Description Unit Definition: One unit incorporates 10 nmol of total

JumpStart REDTaq DNA Polymerase is Sigma’s high deoxyribonucleoside-triphosphates into acid
performance Taq DNA Polymerase blended with precipitable DNA in 30 min at 74 °C.
JumpStart Taq antibody and an inert red dye tracer.
Extensive testing with a variety of primers and Reagents provided
templates indicates that the performance of JumpStart • JumpStart REDTaq DNA Polymerase, Catalog
REDTaq DNA Polymerase is equivalent to, or better Number D0563
than, that of standard Taq polymerase. 1 unit/µL in 20 mM Tris-HCl, pH 8.0, 100 mM KCl,
0.1 mM EDTA, 1 mM DTT, stabilizers, inert dye,
• JumpStart REDTaq DNA Polymerase is the ideal 50% glycerol. Provided as 50, 250 or 2,500 units
enzyme for high throughput and/or multiplex PCR (10 × 250 units)
applications.
• Reactions are prepared the same way as standard • 10× PCR Buffer, Catalog Number P2192
PCR mixtures requiring no additional reaction 100 mM Tris-HCl, pH 8.3 at 25 °C, 500 mM KCl,
preparation steps or protocol changes. 15 mM MgCl2, 0.01% (w/v) gelatin. Provided in
• The hot start mechanism using JumpStart Taq 1.5 ml vials.
antibody prevents non-specific product formation
and allows assembled PCR reactions to be placed Reagents and equipment required but not
at room temperature up to 2 hours without provided
compromising performance.
• Red tracer means quick recognition of reactions to • Primers
which enzyme has been added, as well as visual • DNA to be amplified
confirmation of complete mixing. • 10 mM dATP, Catalog Number D6920
• The enzyme is provided at 1 unit/µL for more • 10 mM dCTP, Catalog Number D7045
accurate volume measurement and less waste. • 10 mM dGTP, Catalog Number D7170
• The enzyme formulation allows aliquots (5-10 µL) • 10 mM TTP, Catalog Number T7791
from the PCR to be directly loaded onto an or
agarose gel without addition of loading buffers. • Deoxynucleotide mix, Catalog Number D7295,
• The red tracer serves as a tracking dye containing 10 mM each dATP, dCTP, dGTP, and
co-migrating at the same rate as a 125 bp TTP
fragment in a 1% agarose gel.
• Water, PCR reagent, Catalog Number W1754
• Mineral oil, Catalog Number M8662 (optional)
Since the red tracer has no effect on the amplification
• 0.5 ml or 0.2 ml thin-walled PCR tubes, Catalog
process, a sample can be easily re-amplified such as
in “nested PCR”. The presence of the dye also has no Numbers P3114 and P3364
effect on automated DNA sequencing; ligase mediated • Thermal cycler
ligations, exonucleolytic PCR product digestion, and
transformation. Though exceptions may exist, the dye
is generally inert in restriction enzyme digestions. If
necessary, the dye can be removed from the amplicon
by routine purification methodologies.
2

Precautions and Disclaimer 1. Add the following reagents to a 0.2 ml or 0.5 ml

This product is for R&D use only, not for drug, PCR tube.
household, or other uses. Please consult the Material
Safety Data Sheet for information regarding hazards Volume Reagent Final
and safe handling practices. Concentration

Storage/Stability
5 µL 10× PCR Buffer 1×
Store at –20 °C.
1 µL* 10 mM dATP 200 µM
JumpStart REDTaq DNA Polymerase, as supplied, will 1 µL* 10 mM dCTP 200 µM
not freeze at –20 °C. It is not recommended to freeze 1 µL* 10 mM dGTP 200 µM
this product at storage temperatures below –20 °C. 1 µL* 10 mM TTP 200 µM
Repeated freeze/thaw cycles may adversely affect its - µL Primers 0.1-0.5 µM
function. 2.5 µL JumpStart REDTaq 0.05 units/µL
DNA Polymerase
Procedure - µL Template DNA 200 pg/µL
Note: The use of DMSO or formamide with JumpStart (typically 10 ng)
REDTaq DNA Polymerase is not recommended due to Water -
interference with the enzyme-antibody complex. Other
co-solvents, solutes (e.g., salts) and extremes in pH or 50 µL Total reaction
other reaction conditions may reduce the affinity of the volume
JumpStart Taq antibody for the Taq DNA polymerase
and thereby compromise its effectiveness. *The individual nucleotides (1 µL of each 10 mM
solution, 4 µL total) may be replaced by 1 µL of
Preparation of PCR Master Mix and Thermal Cycling Deoxynucleotide Mix, Catalog Number D7295.
Parameters
2. Mix gently and briefly centrifuge to collect all
Because Taq DNA polymerase is a magnesium ion- solution at the bottom of the tube.
dependent enzyme, the optimal conditions for the
concentration of Taq, template DNA, primers, and 3. Add 50 µL of mineral oil to the top of each tube to
MgCl2 will depend on the system being utilized. It may prevent evaporation (optional, depending on
be necessary to determine the optimal conditions for model of thermal cycler).
each individual component. This is especially true for
JumpStart REDTaq, cycling parameters, and the 4. Amplification parameters will vary depending on
MgCl2 concentration. It is recommended the enzyme the primers and the thermal cycler used. It may be
and the MgCl2 be titrated to determine the optimal necessary to optimize the system for individual
efficiency. primers, template, and thermal cycler.
To minimize tube-to-tube variation, preparation of a Typical cycling parameters:
PCR master mix with JumpStart REDTaq DNA
Polymerase is recommended. The amount prepared Initial denaturation 94 °C for 1 min
should be based on the number of PCR reactions to
be performed. 25-35 cycles:
Denaturation 94 °C for 30 sec
Annealing 55 °C to 68 °C for 30 sec
Extension 72 °C for 1 min (minimum)*
Final extension: 72 °C for 1 min (minimum)*
Hold 4 °C

* 1 minute minimum or 1 minute per kb expected

amplicon.
 3

5. The amplified DNA can be evaluated by loading Note: A minimum of 1.5 units of JumpStart REDTaq
5-10 µL of the PCR reaction directly onto agarose DNA polymerase must be added per 50 µl reaction to
gel. It is not necessary to add a separate loading ensure enough glycerol is present for direct gel
buffer/tracking dye. loading. The red tracer comigrates with 125 bp
fragment in a 1% agarose gel.

Troubleshooting Guide

Problem Suggestion
No reduction of non-specific Test the PCR system using a manual hot start method.
products is observed when
The use of DMSO or formamide with JumpStart REDTaq DNA Polymerase is
using JumpStart REDTaq DNA
not recommended due to interference with the enzyme-antibody complex. Other
Polymerase.
co-solvents, solutes (e.g., salts) and extremes in pH or other reaction conditions
may reduce the affinity of the JumpStart Taq antibody for the Taq polymerase
and thereby compromise its effectiveness.
Both the JumpStart REDTaq Raise the annealing temperature in 2-3 °C increments. Raising the temperature
PCR and the manual hot start improves the specificity of binding by the primers; however, it may result in
PCR yield multiple nonspecific reduced binding and extension of the primers. If raising the annealing
products. temperature causes a reduced yield of the specific product with only a
proportional reduction of side reaction products, it may be necessary to
redesign the primers.1
Take special precautions to avoid crossover contamination of PCR reactions
with both specific and nonspecific PCR products, including primer-dimer
artifacts.2
The JumpStart REDTaq PCR Titration of JumpStart REDTaq may be necessary to achieve the same degree
yields more non-specific of improvement as with a conventional hot start. This is especially true if the
products than conventional hot PCR reaction conditions vary from those described in this document. In this
start PCR. case, start with a working solution that has a two- to four-fold higher
concentration of JumpStart REDTaq than recommended.
The yield of specific product is Increase the reaction volume to 150 µL or more.
low using JumpStart REDTaq.
Increase the number of amplification cycles. If currently using 25-30 cycles,
increase the cycle number to 35-40. This should increase yields without
significantly increasing side reaction products.

Modify the reaction conditions and/or selection of PCR targets to obtain greater
opportunities for PCR priming. For example, increase the denaturation time up
to 1-1.5 minutes and/or increase the denaturation temperature to as high as
95 °C to overcome denaturation difficulties.
The use of DMSO or formamide with JumpStart REDTaq is not recommended
due to interference with the enzyme-antibody complex.
4

References JumpStart is a trademark of Sigma-Aldrich Co. LLC

1. Huang, L. M., and Jeang, K.-T., BioTechniques REDTaq is a registered trademark of Sigma-Aldrich
16:242-246 (1994) Co. LLC
2. Kwok, S., and Higuchi, R., Nature 339:237-238
(1989)
Related Products

General References Reagents

• Lambda DNA Hind III Digest, Catalog No. D9780
• Griffin, H. G., and Griffin, A. M., (Eds.), PCR
• Enhanced Avian HS RT-PCR kits, Catalog No HSRT100
Technology: Current Innovations, CRC Press, (100 reactions).
1994. Equipment
• Innis, M. A., et al., (Eds.), PCR Strategies, • PCR Multiwell Plate, 96-well, Catalog No. Z374903
Academic Press, New York (1995). • PCR Multiwell Plate, 384-well, Catalog No. Z374911
• Innis, M., et al., (Eds.), PCR Protocols: A Guide to • PCR Microtubes, 0.2 ml, attached caps, Catalog No.
Z374873
Methods and Applications, Academic Press, San
• PCR Microtubes, 0.2 ml strip tubes with strip caps, Catalog
Diego, California (1990). No. Z374962
• Newton, C.R., (Ed.), PCR: Essential Data, John • Sealing accessory for PCR vessels, Micro Mats, Catalog
Wiley & Sons, New York (1995). No. Z374938
• Sambrook, J. F., et al., Molecular Cloning: A • PCR Workstation, 120V, Catalog No. Z376213
Laboratory Manual, Third Edition, Cold Spring • PCR Workstation, 240V, Catalog No. Z376221
Harbor Laboratory Press, New York (2000),
Catalog Number M8265. NOTICE TO PURCHASER: LIMITED LICENSE

Use of this product is covered by one or more of the

following US patents and corresponding patent claims
outside the US: US 8,404,464 and US 7,972,828. The
purchase of this product includes a limited, non-transferable
immunity from suit under the foregoing patent claims.
.

GAR,PHC 12/13-1

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