Primer/Probe Set for Housekeeping Gene in Human Sinus Tissue

Northwestern University
Medecine
Graduate Student

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Company:

Thermo Fisher (Applied Biosystems)

Product Name:

Primer/probe set for beta-glucuronidase

Catalog Number:

4326320E

We work with human sinus tissue and usual housekeeping genes for RT-PCR, like GAPDH, were found to have different baseline expression in nasal polyps compared to uncinate tissue, which could confound results. We found that beta-glucuronidase, or bGUS, had the most consistent expression in nasal polyps and uncinate tissue.

Experimental Design and Results Summary

Application

RT-PCR

Starting Material

Human sinus tissue lysates

Protocol Overview

The primer/probe set is a 20x solution, so it you're using 20uL reactions you only need 1uL/reaction.

Tips

This is available on VIC, so you can multiplex with FAM primer probe sets to save sample and reagents

Results Summary

bGUS is the most reliable housekeeping gene for human sinus tissue.

Additional Notes

Oncostatin M promotes mucosal epithelial barrier dysfunction, and its expression is increased in patients with eosinophilic mucosal disease. Pothoven KL, Norton JE, Hulse KE, Suh LA, Carter RG, Rocci E, Harris KE, Shintani-Smith S, Conley DB, Chandra RK, Liu MC, Kato A, Gonsalves N, Grammer LC 3rd, Peters AT, Kern RC, Bryce PJ, Tan BK, Schleimer RP. J Allergy Clin Immunol. 2015 Sep;136(3):737-746.e4. doi: 10.1016/j.jaci.2015.01.043.

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Summary

The Good

Great primer/probe set, super easy

The Bad

More expensive than sequencing primers and probes

The Bottom Line

This is the best housekeeping gene for human sinus tissue, and multiplexing with other genes saves reagents and sample.

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