Megaplex™ PreAmp Primers, Rodent Pool Set v3.0

For use when assay sensitivity is of the utmost importance, Megaplex™ PreAmp Primers significantly enhance the ability to detect lowRead more

Catalog Number	Quantity
4444747	50 reactions

Catalog number 4444747

Price (USD)

Quantity:

50 reactions

For use when assay sensitivity is 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, Rodent Pools Set v3.0 is used with Megaplex™ RT Primers, Rodent Pools 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® MicroRNA Arrays.

For Research Use Only. Not for use in diagnostic procedures.

Specifications

No. of Reactions50 Reactions

PrimerPreAmp Primers

Product LineMegaplex™, TaqMan™

Product TypePreamp Primer

Quantity50 reactions

SpeciesRat, Mouse

Sufficient For50 Reactions

GC-Rich PCR PerformanceLow

PCR MethodqPCR

Unit SizeEach

Contents & Storage

Store at -20°C.

Generating response

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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.

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