FastStart Taq DNA Polymerase
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for "Hot Start" PCR amplification
of up to 3 kb RNA targets Cat. No. 2 158 264
50 units for 25 PCR reactions
Cat. No. 2 032 902 100 units for 50 PCR reactions
Cat. No. 2 032 929 2× 250 units for 250 PCR reactions
Cat. No. 2 032 937 4× 250 units for 500 PCR reactions
Cat. No. 2 032 945 10× 250 units for 1250 PCR reactions
Cat. No. 2 032 953 20× 250 units for 2500 PCR reactions
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| Description |
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FastStart Taq DNA Polymerase has been developed by Roche to make Polymerase
Chain Reaction (PCR) more specific and sensitive in a convenient and rapid
way. With FastStart Taq DNA Polymerase, "Hot Start" PCR can
be applied for genomic DNA and cDNA templates, eliminating the extra handling
steps or additional time required associated with up to now known "Hot
Start" methods.
FastStart Taq DNA Polymerase is a thermostable, modified form of recombinant
Taq DNA Polymerase. It is inactive at temperatures below 75°C, but can
be activated by a 4 min 95°C incubation step. The combination of FastStart
Taq DNA Polymerase and the optimized PCR buffer minimizes non-specific
amplification products and primer dimers allowing highest sensitivity.
The provided GC-RICH solution, a PCR additive that facilitates amplification
of difficult templates by modifying the melting behavior, will improve
PCR performance on templates rich in secondary structures or GC content.
For further information please download the FastStart
product flyer (950 Kbytes PDF file). Please note that this file is
in Acrobat
PDF format and must be viewed, searched, and printed with version
3.0 or higher of the free Adobe
Acrobat Reader.
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| Application |
FastStart Taq DNA Polymerase is an ideal tool
for "Hot Start" PCR, because the enzyme remains inactive during
PCR set-up and prior to the initial denaturation step. Since it is inactive
at low temperatures, FastStart Taq DNA Polymerase cannot elongate non-specific
primer-template hybrids that may form at those temperatures. FastStart Taq
DNA Polymerase delivers superior results if used for:
- Amplification of genomic DNA and cDNA targets up to 3 kb long with
high specificity, sensitivity, and yield
- Multiplex PCR
- Difficult templates e.g. secondary structures or GC-rich sequences
- Carry-over prevention
- Automated PCR e.g. handling at room temperatures
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| Experimental results |
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Figure 1: Specificity and sensitivity comparison in PCR using commercially
available hot start systems. |
| Varying amounts of human genomic DNA were used
for the amplification of a single 130 bp fragment from the tissue plasminogen
activator (tPA) gene. Manufacturers' recommended initial product activation
times were used when applicable. Cycling conditions: 35 cycles (95°C / 30
seconds - 60°C / 30 seconds - 72°C / 60 seconds) final extension at
72°C for 7 minutes. A: FastStart Polymerase; B: Taq DNA Polymerase; C: Supplier
A, modified hot start polymerase, buffer I; D: Supplier A, modified hot
start polymerase buffer II; E: Taq DNA Polymerase with anti-Taq antibody;
F: Supplier B, antibody-modified hot start polymerase. |
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Figure 2: Sensitivity comparison when amplifying a single-copy gene. |
| Varying amounts of human genomic DNA were used
for amplification of a single 375 bp fragment from the tissue plasminogen
activator (tPA) gene. Manufacturers' recommended initial product-activation
times were used when applicable. Cycling conditions: 40 cycles (95°C / 30
seconds - 55°C / 30 seconds - 72°C / 60 seconds) final extension at 72°C
for 7 minutes. A: FastStart Polymerase; B: Taq DNA Polymerase; C: Supplier
A, modified hot start polymerase, buffer I; D: Supplier A, modified hot
start polymerase buffer II; E: Taq DNA Polymerase with anti-Taq antibody;
F: Supplier B, antibody-modified hot start polymerase. |
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| Key advantages |
- Minimize optimization difficulties in all types of PCR with FastStart
Polymerase’s specially designed and optimized buffers.
- Achieve results with difficult templates using the optimized GC-RICH
Resolution Solution
- Amplify templates up to 3 kb and even difficult templates (e.g.,
high GC content, secondary structure) better than other hot start systems
- Produce results across a wide range of magnesium concentrations
- Eliminate wax barriers, beads, hot start antibodies, manual hot start,
and the need to set up PCR reactions on ic
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