Quantification of Gene Expression using Mastercycler ep gradient

University of Nebraska Medical Center
Biochemistry and Molecular Biology
Postdoctoral Research Associate

Overall

Performance

Ease-of-Optimization

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Company:

Eppendorf

Product Name:

Mastercycler ep gradient

Catalog Number:

5341

Our lab is investigating quantitative expression of various genes under different experimental conditions. We routinely employ qPCR for the analysis of different gene expression in both normal and cancerous cells before and after various drug treatments. We used Master cycler epgradient machine from Eppendorf for this purpose and obtained consistent result all the time.

Experimental Design and Results Summary

Application

Quantitative gene expression

Starting Material

cDNA synthesized from mRNA obtained from human cells.

Protocol Overview

Initially, mRNA was obtained from human cells and converted into cDNA. cDNA was loaded on to 96 well plate along with forward and reverse primers. Syber green (5 ul) was added and final volume was adjusted to 10 ul with mol bio grade water. Plate was centrifuged and loaded into the machine for analysis.

Tips

We used 2 ug of mRNA for the synthesis of cDNA and from this preparation, 2 ul of cDNA is used for the analysis.

Results Summary

We obtained high quality results for the expression of genes uder various conditions. The results were reproducible and thus we could able to publish these results in peer-reviewed journals.

Features Summary

Simple and user-friendly

Additional Notes

None

Image Gallery

Summary

The Good

Software is user friendly

The Bad

None

The Bottom Line

Over all, the equipment is very help full in determining the expression of genes in different samples with very limited small amount of starting material.

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