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

AgPath-ID™ One-Step RT-PCR Reagents

Los reactivos para RT-PCR AgPath-ID™ de un paso se han diseñado para una amplificación sensible y sólida de dianas de ARN mediante una rápida estrategia de PCR de transcripción inversa en tiempo real (RT-PCR) TaqMan™ de tubo único.
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Número de catálogoN.º de reacciones
AM1005100 reacciones
4387424500 reacciones
43873911000 reacciones
Número de catálogo AM1005
Precio (MXN)
6,187.14
Each
Añadir al carro de la compra
N.º de reacciones:
100 reacciones
Precio (MXN)
6,187.14
Each
Añadir al carro de la compra

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.
Especificaciones
DescripciónFunciona en todas las plataformas AB, pero está optimizado para la 7500.
Fidelidad (frente a Taq)25 X
Para utilizar con (equipo)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
FormatoTubo
N.º de reacciones100 reacciones
Colorante de referencia pasivaROX (premezclado)
PolimerasaADN polimerasa AmpliTaq Gold
Línea de productosAgPath-ID, Ambion
Tipo de productoOne-Step RT-PCR Reagent
Cantidad100 reactions
Transcriptasa inversaArrayScript™
Tipo de muestraARN
Condiciones de envíoHielo seco
Suficiente para100 reacciones
Volumen1.75 mL
Concentración25X
Método de detecciónSonda de cebado
GC-Rich PCR PerformanceHigh
Método de PCRQRT-PCR de 1 paso
Velocidad de reacciónEstándar
Unit SizeEach
Contenido y almacenamiento
Suficiente para 100 reacciones de 25 μl, contiene:
• 1375 μl 2X tampón para RT-PCR
• 110 μl 25X mezcla de enzimas para RT-PCR
• 1,75 ml de agua sin nucleasas.

Preguntas frecuentes

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.

View all AgPath-ID™ One-Step RT-PCR Reagents FAQs