Precise Viral RNA Isolation from supernatant of retroviral producer cell lines

University of Michigan
Pharmaceutical Sciences
Research Fellow

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Company:

Qiagen

Product Name:

QIAamp Viral RNA Mini Kit

Catalog Number:

52904

Producer cell lines to obtain retroviruses and adenoviruses are commonly used in most viral therapeutics and transduction labs. In order to precisely quantify the number of viral particles, efficient viral RNA isolation is required which was easily done via this kit.

Experimental Design and Results Summary

Application

Quantification of viral particles

Starting Material

Cell supernatant from producer cell lines - See http://www.clontech.com/US/Products/Viral_Transduction/Retroviral_Packaging/High-Titer_Retrovirus

Protocol Overview

Protocol suggested by the manufacturer was used. No other optimization was done.

Tips

Make sure lab bench is extremely clean to avoid other cellular contamination

Results Summary

Isolation was a breeze with this kit. The attached image shows the viral rna isolated following reverse transcription, traditional pcr, and gel electrophoresis. Lane 1 shows the ladder and Lanes 2 and 3 show our cDNA for the RNA which we were interested in. For reverse transcription, we used the following kit - http://www.lifetechnologies.com/order/catalog/product/N8080234

Additional Notes

None

Image Gallery

Summary

The Good

Quick and easy

The Bad

None

The Bottom Line

Highly repeatable and precise RNA isolation of viral particles

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