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End-Point RT-PCR
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Applications
- Highly accurate RT-PCR amplification of RNA templates for cloning, sequencing,
expression, and transcription analysis.
- All-purpose RT-PCR in which full-length amplification and high accuracy
are essential.
- Multiplex RT-PCR.
The MasterAmp™ High Fidelity RT-PCR Kit is ideal for synthesis of
accurate double-stranded cDNA products that will be used for cloning, sequencing,
expression, or transcription analysis. The kit uses MMLV-RT Plus, a reverse
transcriptase with an unsurpassed accuracy and reliability, and MasterAmp
TAQurate™ DNA Polymerase, a high-fidelity PCR enzyme mix (Fig. 1).
The kit also includes an optimized 2X RT-PCR Buffer PreMix (containing dNTPs,
MgCl2, and buffer), random and oligo(dT) primers, and the MasterAmp
PCR Enhancer with betaine,* which can substantially improve the yield and
specificity of RT-PCR amplification.
Benefits
- High-fidelity RT-PCR (i.e., low error rates).
- Amplification of full-length products from RNA templates up to ~6 kb
(Fig. 2).
- Random and oligo(dT) primers included for first-strand cDNA synthesis.
- Unique PreMix reduces pipetting steps and chances of contamination.
- Uses MasterAmp PCR Enhancer for better amplification of high-GC or other
difficult templates.
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Figure 1. Different RT-PCR protocols that can be used with the MasterAmp™ High
Fidelity RT-PCR Kit. |
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Figure 2. The MasterAmp™ High Fidelity RT-PCR Kit amplifies full-length
products from RNA templates up to ~6 kb. RT-PCR products from Tobacco
Mosaic Virus RNA with primers that amplify a 5.5-kb region of the viral RNA.
Lane M, Kilobase ladder; lane 1, RT-PCR using the MasterAmp one-step protocol;
lane 2, RT-PCR using MMLV-RT and Taq DNA polymerase with a standard
two-step procedure; lane 3, RT-PCR using Tth DNA polymerase. |
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Figure 3. Amplification of a variety of RNA templates using a one-step
protocol with the MasterAmp™ High Fidelity RT-PCR Kit. Lane M,
100-bp ladder; lane 1, human lung β-actin; lane 2, human lung GAPDH;
lane 3, region of TMV; lane 4, E. coli 16S rRNA; lane 5, human CGα;
lane 6, human heart β-actin; lane 7, region of TMV. |
*Covered by issued and/or pending patents.
MasterAmp™ High Fidelity RT-PCR Kit. Purchase of this product includes
an immunity from suit under patents specified in the product insert to use
only the amount purchased for the purchaser’s own internal research.
No other patent rights (such as 5´ Nuclease Process patent rights) are
conveyed expressly, by implication, or by estoppel. Further information on
purchasing licenses may be obtained by contacting the Director of Licensing,
Applied Biosystems, 850 Lincoln Centre Drive, Foster City, California 94404,
USA.
CopyControl™ cDNA, Gene and PCR Cloning Kits
FailSafe™ GREEN and PROBES, TAQurate™ GREEN and PROBES, MasterAmp™ GREEN, TAQXpedite™ GREEN (discontinued)
FailSafe™ PCR System
MasterAmp™ RT-PCR Kit for High Sensitivity
MasterPure™ Complete DNA and RNA Purification Kit
MonsterScript™ 1st-Strand cDNA Synthesis Kit
MonsterScript™ Reverse Transcriptase
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