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

Megaplex™ PreAmp Primers, Human Pool Set v3.0

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For use when assay sensitivity of the utmost importance, Megaplex™ PreAmp Primers significantly enhance the ability to detect low expressedRead more
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Catalog number 4444748
Price (USD)
1,308.00
Each
Add to cart
Price (USD)
1,308.00
Each
Add to cart
For use when assay sensitivity of the utmost importance, Megaplex™ PreAmp Primers significantly enhance the ability to detect low expressed microRNAs enabling the generation of a comprehensive expression profile using far less sample, as low as 1 ng of input total RNA. With content matched to the Megaplex™ RT Primers, Megaplex™ PreAmp Primers, Human Pool Set v3.0 is used with Megaplex™ RT Primers, Human Pool Set v3.0. Following reverse transcription using Megaplex™ RT Primers, Megaplex™ PreAmp Primers along with the TaqMan® PreAmp Master Mix uniformly amplify all microRNAs prior to quantitation by real-time PCR using TaqMan® Array MicroRNA Cards.
For Research Use Only. Not for use in diagnostic procedures.
Specifications
GC-Rich PCR PerformanceLow
PCR MethodqPCR
Green FeaturesSustainable packaging
PrimerPreAmp Primers
Product LineMegaplex™
Product TypePreamp Primer
SpeciesHuman
Sufficient For50 Reactions
No. of Reactions50 reactions
Unit SizeEach
Contents & Storage
Store at -20°C

Frequently asked questions (FAQs)

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.

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