AgPath-ID™ One-Step RT-PCR Reagents
AgPath-ID™ One-Step RT-PCR Reagents
Applied Biosystems™

AgPath-ID™ One-Step RT-PCR Reagents

AgPath-ID™ One-Step RT-PCR Reagents are designed for sensitive, robust amplification of RNA targets using a rapid single-tube TaqMan™ real-time reverse transcription PCR (RT-PCR) strategy.
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Catalog NumberNo. of Reactions
AM1005100 Reactions
4387424500 Reactions
43873911000 Reactions
Catalog number AM1005
Price (CNY)
2,813.00
Each
Add to cart
No. of Reactions:
100 Reactions
Price (CNY)
2,813.00
Each
Add to cart

AgPath-ID™ One-Step RT-PCR Reagents are designed for sensitive, robust amplification of RNA targets using a rapid single-tube TaqMan™ real-time reverse transcription PCR (RT-PCR) strategy.

  • Recommended for RNA pathogen amplification
  • Consistent amplification of RNA targets with high specificity and sensitivity
  • Optimized to work with your target-specific primers and probes
  • Contains ROX for quantitative fluorescent signal normalization

AgPath-ID™ One-Step RT-PCR Reagents are configured for fast and simple reaction setup. The reactions are assembled in a single tube, minimizing sample handling errors and expediting set-up time. Once assembled, results are available in approximately one hour. The 25X RT-PCR Enzyme Mix included in the kit contains highly efficient ArrayScript™ Reverse Transcriptase, a mutant MMLV RT that produces high cDNA yields, and AmpliTaq Gold™ polymerase, the preferred hot-start DNA polymerase for specific target amplification. The 2X RT-PCR Buffer has been optimized for efficient, robust reverse transcription and PCR includes the passive reference dye, ROX™ Dye, for quantitative fluorescent signal normalization.

For Research Use Only.
Specifications
DescriptionWorks on all AB platforms, but is optimized for the 7500.
Fidelity (vs. Taq)25 X
For Use With (Equipment)7500 System, 7500 Fast System, 7900HT System, StepOne™, StepOnePlus™, ViiA™ 7 System, QuantStudio™ 3, QuantStudio™ 5, QuantStudio™ 6 Flex, QuantStudio™ 7 Flex, QuantStudio 6 Pro, QuantStudio 7 Pro, QuantStudio™ 12k Flex, QuantStudio™ Absolute Q Digital PCR System
FormatTube
No. of Reactions100 Reactions
Passive Reference DyeROX (Pre-mixed)
PolymeraseAmpliTaq Gold DNA Polymerase
Product LineAgPath-ID™, Ambion™
Product TypeOne-Step RT-PCR Reagent
Quantity100 reactions
Reverse TranscriptaseArrayScript™
Sample TypeRNA
Shipping ConditionDry Ice
Sufficient For100 Reactions
Volume1.75 mL
Concentration25X
Detection MethodPrimer-probe
GC-Rich PCR PerformanceHigh
PCR Method1-step RT-qPCR
Reaction SpeedStandard
Unit SizeEach
Contents & Storage
Sufficient for 100 reactions of 25 μL, contains:
• 1375μL 2X RT-PCR Buffer
• 110μL 25X RT-PCR Enzyme Mix
• 1.75mL Nuclease-free Water.

Frequently asked questions (FAQs)

With the AgPath-ID One-Step RT-PCR Reagents, I am getting signal detection in the no-template control (NTC) reaction. Why is this?

This is likely due to PCR contamination. Here are some recommendations:

- Repeat the qRT-PCR reaction with fresh reagents and decontaminated pipettors.
- Set up and run the qRT-PCR reaction in an area that is isolated from areas used for nucleic acid isolation and PCR product analysis.
- The Reverse Transcriptase enzyme contained in this kit is produced using an E. coli expression vector containing a proprietary version of the MMLV pol gene (GenBank accession no. J02255) expressed from pET-24(+). It is possible that a minimal amount of the expression vector could be carried over into the final mastermix formulation. If you are targeting MMLV, a related virus, or any of the plasmid sequence, we recommend designing primer sequences for target sequences not contained in the expression vector.

What can I do to improve the sensitivity of my qPCR assay?

If you are targeting a low-abundance gene, you may have trouble getting Ct values in a good, reliable range (Ct > 32). To increase the sensitivity of the assay, you may want to consider the following:

- Increase the amount of RNA input into your reverse transcription reaction, if possible
- Increase the amount of cDNA in your qPCR reaction (20% by volume max)
- Try a different reverse transcription kit, such as our SuperScript VILO Master Mix, for the highest cDNA yield possible
- Consider trying a one-step or Cells-to-CT type workflow (depending on your sample type)

How do I set the baseline for my qPCR experiment?

Most times your instrument software can automatically set a proper baseline for your data. Check out our short video, Understanding Baselines, for more information on how to set them (https://www.youtube.com/watch?feature=player_embedded&v=5BjFAJHW-bE).

How do I set the threshold for my qPCR experiment?

In most cases your instrument software can automatically set a proper threshold for your data. Check out our short video, Understanding Thresholds, for more information on how to set them (https://www.youtube.com/watch?feature=player_embedded&v=H_xsuRQIM9M).

I am not getting any amplification with my TaqMan Assay or SYBR Green primer set. What is causing this?

There could be several reasons for no amplification from an assay or primer set. Please see these examples and suggested solutions in our Real-Time Troubleshooting Tool (https://www.thermofisher.com/us/en/home/life-science/pcr/real-time-pcr/qpcr-education/real-time-pcr-troubleshooting-tool/gene-expression-quantitation-troubleshooting/no-amplification.html) for more details.

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