Megaplex™ PreAmp Primers, Rodent Pool Set v3.0

Para su uso cuando la sensibilidad del ensayo es de la máxima importancia, los cebadores Megaplex™ PreAmp mejoran significativamente laMás información

Número de catálogo	Cantidad
4444747	50 reacciones

Número de catálogo 4444747

Precio (USD)

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 contenido adaptado a los cebadores Megaplex™ RT, los cebadores Megaplex™ PreAmp, el conjunto de grupos de roedores v3.0 se utiliza con cebadores Megaplex™ RT, conjunto de grupos de roedores 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 matrices de MicroARN de TaqMan®.

Para uso exclusivo en investigación. No apto para uso en procedimientos diagnósticos.

Especificaciones

N.º de reacciones50 reacciones

CebadorCebadores PreAmp

Línea de productosMegaplex, TaqMan

Tipo de productoCebador PreAmp

Cantidad50 reacciones

EspecieRata, ratón

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.

View all Megaplex™ PreAmp Primers, Rodent Pool Set v3.0 FAQs