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Oral Biology pp 299-306 | Cite as

A Protocol for the Determination of the Methylation Status of Gingival Tissue DNA at Specific CpG Islands

  • Trudy J. MilneEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1537)

Abstract

Tissue biopsies are very precious. A method that allows the isolation of a high quality and quantity of genomic DNA, total RNA, and total protein from a single biopsy that is suitable for downstream applications (e.g., DNA methylation analysis, quantitative PCR, and gel electrophoresis techniques) is very desirable. The method described here combines a tissue stabilization reagent combined with a spin-column method for the simultaneous purification of gingival tissue DNA, RNA, and protein. The genomic DNA is then used for quantitative analysis of DNA methylation using real-time PCR (the qAMP method). Subsequent data analysis is very straightforward using online software. Future analyses may include the RNA transcript analysis as well as protein expression levels of genes identified as differentially methylated.

Key words

qAMP Regional DNA methylation Gingival tissue Genomic DNA Total RNA Allprotect™ tissue reagent AllPrep® DNA/RNA/protein kit 

Notes

Acknowledgments

This work was supported by a grant from the New Zealand Dental Association.

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Copyright information

© Springer Science+Business Media LLC 2017

Authors and Affiliations

  1. 1.Faculty of Dentistry, Sir John Walsh Research InstituteUniversity of OtagoDuendinNew Zealand

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