SPAM-LOW: [Histonet] glutaraldehyde fixed tissues and genomic DNA

[Patsy Ruegg]

Wow this sounds like a molecular biology question that will have to be
figured out and probably no one has done that yet for this particular
situation.  Please write it up if you do figure it out.

Regards,
Patsy

Patsy Ruegg, HT(ASCP)QIHC
IHCtech
12635 Montview Blvd. Ste.215
Aurora, CO 80045
720-859-4060
fax 720-859-4110
www.ihctech.net 
www.ihcrg.org


-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of
ejschmid <@t> ucalgary.ca
Sent: Monday, November 15, 2010 11:01 AM
To: histonet <@t> lists.utsouthwestern.edu
Subject: SPAM-LOW: [Histonet] glutaraldehyde fixed tissues and genomic DNA

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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