[Histonet] glutaraldehyde fixed tissues and genomic DNA

ejschmid <@t> ucalgary.ca ejschmid <@t> ucalgary.ca
Mon Nov 15 12:01:23 CST 2010


Hello,

I have PFA+glutaraldehyde fixed tissues that I might want to digest down
in order to extract genomic DNA suitable for bisulfite sequencing.

The tissues are embryonic mouse heads. The fixative is a PFA +
glutaraldehyde mix:  3.6 PFA with 5% glut. The glut is grade 1 or electron
miccroscopy grade.

I'd like to pro-K the tissues and then phenol-chloroform extract genomic DNA.

My hope is that the amount and quality of the DNA will be suitable for
bisulfite conversions for methylation anaylsis. Will the DNA be abundant?
Will it be fairly protein free (does the fixative fix protein to the
DNA?).

Thanks

Eric
University of Calgary
Facutly of Medicine




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