Real-time PCR Instrument Performs Below Other Brands

May 29, 2014

Overall

Performance

Ease-of-Optimization

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Company:

Eppendorf

Product Name:

Mastercycler Realplex Ep Real-Time PCR System

Catalog Number:

97060-540

We perform gene expression analysis by RT-qPCR for a wide range of mRNA and miRNA targets by absolute and relative quantification.

Experimental Design and Results Summary

Application

Quantification of gene expression

Starting Material

Mammalian cell cultures

Protocol Overview

Starting material prepared using standard protocols:cDNA: RNA extracted and gDNA removed using the Maxwell automated RNA extraction system (simplyRNA LEV kit). 500ng of RNA converted to cDNA using the High Capacity cDNA synthesis kit (Life Technologies). qPCR performed using plastics from USA scientific and mastermix from Kappa Bioscience.

Tips

NA

Results Summary

The mastercycler is capable of detecting cDNA by qPCR over a wide range of input values.

Features Summary

NA

Additional Notes

None

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Summary

The Good

The product does what it is intended to do.

The Bad

The analysis software is very poor. Background calculations generally require manual adjustment after every run. We have encountered multiple technical probems with this system. Detection in the HEX channel is very poor on our instrument. When performing melt analysis we frequently see jumps in the signal detection which are clearly artefacts as a result of detector failures. The instrument has been returned to fix these problems twice but the problems persist. Manufacturer did not provide a loan unit while the instrument was down and repairs took 5 weeks.

The Bottom Line

We don't suggest to use this cycler. The Applied Biosystems StepOne Plus and Roche Lightcycler 96 perform in a superior way.

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