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Biology

Stimulation of Cytoplasmic DNA Sensing Pathways In Vitro and In Vivo

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JoVE Journal Biology

Stimulation of Cytoplasmic DNA Sensing Pathways In Vitro and In Vivo

Stimulation of Cytoplasmic DNA Sensing Pathways In Vitro and In Vivo

DOI:

10.3791/51593-v

•

11:44 min

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September 18, 2014

•

Chih Hung Ku, Brian J. Ferguson

¹Department of Pathology,University of Cambridge

Chapters

00:05Title
01:17Concatemerization of Immuno-stimulatory DNA Sequences
02:43DNA Purification and Analysis
05:55Transfection of dsDNA Concatemers In Vitro
06:47Transfection of dsDNA Concatemers In Vivo
08:29Processing of DNA Stimulated Samples and Quantitative PCR Analysis
10:32Results: Concatermerized DNA Stimulate an Innate Immune Response In Vitro and In Vivo
11:14Conclusion

Summary

Automatic Translation

The aim of the protocol is to use optimal methods for stimulating the cytoplasmic DNA sensing pathways in cells and in vivo. This is achieved by improving the generation of long, double-stranded DNA during blunt-end ligation. Cells or mice are then transfected using a lipid transfection reagent.

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