Great Real-Time Machine from a Trusted Company

June 12, 2014

Overall

Performance

Ease-of-Optimization

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University of California San Diego (UCSD)
Medicine
Post-doctoral researcher

Company:

Life Technologies/Applied Biosystems

Product Name:

StepOne Plus Real-Time PCR

Catalog Number:

4376600

I use real-time PCR machine for various purposes, mainly for checking for the knockdown/over expression of various genes after transfection and to quantify the signals for ChIP experiments. We chose this machine for real-time because of the reputation of the company and because of the reproducibility and time saving features. Having StepOne software for data analysis was also helpful.

Experimental Design and Results Summary

Application

Chromatin immunoprecipitation (ChIP)

Starting Material

Sonicated chromatin from formaldehyde cross-linked HeLa cells

Protocol Overview

ChIP was performed with antibodies against FLAG and myc tag after over-expression of plasmid containing both tags.

Tips

One should be careful to orient the pat in the right direction and that the surface of the PCR plate is clean of any dust particles.

Results Summary

Real-time quantitative PCR was performed using SYBR green from Bio-Rad. The cT values were taken after the completion of the experiment which were then used to calculate the fold enrichment for the antibodies used over IgG.

Features Summary

Touch-screen, easy to use intuitive interface, SteoOne software for analysis, short experimental duration

Additional Notes

None

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Summary

The Good

Easy to use and simplified protocol, touch screen, easy reaction set-up, reproducibility, time saving

The Bad

StepOne software is Windows only (will not work on Windows 8). Mac users cannot use it at all.

The Bottom Line

Step One plus is an ideal instrument for real-time PCR. The results are consistent and reproducible. StepOne plus can handle four colors at once, giving you flexibility of multiplexing.

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