Megaplex™ PreAmp Primers, Human Pool B v3.0
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Applied Biosystems™

Megaplex™ PreAmp Primers, Human Pool B v3.0

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Para su uso cuando la sensibilidad del ensayo es de la máxima importancia, los cebadores Megaplex™ PreAmp mejoran significativamente laMás información
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Número de catálogoCantidad
444430350 reacciones
Número de catálogo 4444303
Precio (MXN)
-
Cantidad:
50 reacciones
Para su uso cuando la sensibilidad del ensayo es de la máxima importancia, los cebadores Megaplex™ PreAmp mejoran significativamente la capacidad de detectar microARN de baja expresión, lo que permite generar un perfil de expresión completo utilizando mucha menos muestra, tan solo 1 ng del ARN total de entrada.Con el contenido adaptado a los cebadores de RT Megaplex™, cebadores de PreAmp Megaplex™, Human Pool B v3.0 está diseñado para usarse con los imprimadores de RT Megaplex™, Human Pool B v3.0.Después de la transcripción inversa utilizando los cebadores Megaplex™ RT, los cebadores Megaplex™ PreAmp junto con la mezcla maestra TaqMan® PreAmp amplifican uniformemente todos los microARN antes de la cuantificación mediante PCR en tiempo real utilizando las tarjetas de MicroARN de matriz TaqMan®.
Para uso exclusivo en investigación. No apto para uso en procedimientos diagnósticos.
Especificaciones
Características ecológicasEmbalaje sostenible
N.º de reacciones50 reacciones
CebadorCebadores PreAmp
Línea de productosMegaplex
Tipo de productoCebador PreAmp
Cantidad50 reacciones
Condiciones de envíoTemperatura ambiente
EspecieHumano
Suficiente para50 reacciones
GC-Rich PCR PerformanceBajo
Método de PCRqPCR
Unit SizeEach
Contenido y almacenamiento
Almacenar a -20°C

Preguntas frecuentes

Can Ct's greater than a cut-off be considered valid results?

Yes they can. However, it is important to recognize the true linearity and detection limits of your assay: Ct values above the cut-off can indicate non-specific amplification, unless your NTC is a true- no-target control, and you have run a statistically significant number of replicates. Any results with Ct above the recommended cut-off need to be validated with individual assays on plates.

What is a good Ct cut-off for the TaqMan MicroRNA Array Cards and TaqMan Advanced miRNA Array Cards? In other words, beyond what Ct should I not trust the data?

The typical Ct cut-off on TaqMan Array Cards is 32, which is equivalent to Ct 35 on a plate (10 µl reaction). Previous studies show that if you use pre-amplification, a Ct cut-off of 29 or 30 can be used to reduce numbers of false positives (see Technical Note Optimized protocols for human or rodent microRNA profiling with precious samples). To ensure that you have selected a correct cut-off, you should run replicates of the same sample and use Ct cut-off before you see an increase in the Standard Deviation.

Why does the negative control well show amplification when doing microRNA analysis using Megaplex Primer Pools and TaqMan Array Cards?

Most likely the reagents or the cDNA template are contaminated. Please follow established PCR laboratory best practices.

Why do I have poor reproducibility across technical replicates when doing microRNA analysis using Megaplex Primer Pools and TaqMan Array Cards?

Most likely the reagents were not adequately mixed. Ensure that all samples and reagents are mixed well.

When doing microRNA analysis using Megaplex Primer Pools and TaqMan Array Cards, why are the Ct values for the endogenous controls highly variable across the sample set?

Highly variable values for endogenous controls is most likely due to biological variation. We recommend that you use an alternative endogenous control, such as a non-variable miRNA.