From john.mcewan agresearch.co.nz Sun Jul 4 04:50:22 2010
From: "McEwan, John" <john.mcewanagresearch.co.nz>
To: Multiple Recipients of <angenmapanimalgenome.org>
Subject: RE: Whatman FTA papers
Date: Sun, 04 Jul 2010 04:50:22 -0500
Dear Miguel
We have used the Gensolve kit on reasonable numbers of blood samples from
sheep collected on FTA for running on Ovine SNP chips. Jerry Taylor's group
have described the method http://www.ncbi.nlm.nih.gov/...MC2704227/
We find with a 6mM diameter punch we get enough for a SNP chip without
amplification and very good results. We have used samples collected 2-5 years
ago that have been stored at room temperature. The great thing about this
extraction method is that it does not denature the DNA and you get sufficient
good quality DNA of reasonable length. If you try it then I suggest contact
the kit manufacturers GenVault direct http://www.genvault.com as they are very
helpful. Its not a "cheap" method but compared to the price of a SNP chip its
trivial. The standard extraction method is alkaline lysis at higher
temperatures okay for running say microsatellites or single SNP PCR based
assays but not much else.
For mammals here are some figures
* A 50ul blood causes a 15mm diameter blot on FTA
* white blood cells are 7000cells/ul (range 4000-12000)
* pgDNA/cell ~5.98
Using the above figures a 2mm diameter punch has ~37ng DNA and a 6mm
diameter punch ~333ng. Normally ~40-60% can be extracted.
Hope this gets you started.
Cheers
John McEwan
-----Original Message-----
.From: Miguel Perez Enciso [mailto:Miguel.Perezuab.cat]
.Sent: Friday, 2 July 2010 11:29 p.m.
.To: Multiple Recipients of
.Subject: Whatman FTA papers
Dear colleaugues, Does anyone have experience with extracting DNA from
blood collected with FTA papers? About how much dna can you recover? I
understand that you get very little DNA and that you require a protocol to
genome wide amplify it (eg, genomiphi). Any help will be welcome,
outstanding ones will be rewarded with beer at Leipzig meeting, (sorry not
going to ISAG). Thanks!
=======================================
Miguel Perez-Enciso
ICREA professor
Dept. Ciencia Animal i dels Aliments
Facultat de Veterinaria
Universitat Autonoma de Barcelona
08193 Bellaterra, SPAIN
Phone: +34 93 581 4225
Fax: +34 93 581 2106
miguel.perezuab.es
=======================================
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