I am interested in extracting DNA from the hearts of P0 mouse in order to genotype the heart (for heart-specific recombinations). I want to embed hearts in gelatin in order to do cryosections of the heart (for further analysis) and leave a portion of the ventricle apex for DNA extraction and genotyping.
I have found a protocol for DNA isolation but it doesn't say whether the tissue is embedded (e.g. gelatin, OCT, etc.) or not. There is a procedure for "un-embedding" tissues from gelatin (i.e. bath at 37 ºC) but I'm not sure whether it'll work out for the DNA isolation.
Thank you in advance!
http://diyhpl.us/~bryan/irc/protocol-online/protocol-cache/dnaisopt.htm
Have a look on this article, hope it is helpful
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC1876866/
Hi Oriol, I'm not sure if this will be helpful to you, but I used to work on fresh-frozen cancer tissues that required OCT embedding and cryosectioning of a sample first to identify areas of high tumour content for DNA isolation. We would then cut out the selected tumour area with a scalpel while the sample was still frozen (could do this on dry ice or LN2) so that it doesn't warm up and trim as much OCT off as possible. Following that, the sample would be disrupted and DNA extracted with a kit - in this case a spin column kit such as the Qiagen Blood and Tissue Kit.
We were using this DNA for high sensitivity downstream applications and didn't have issues relating to OCT contamination that I know of.
Good luck.
Even if you can't remove gelatin completely from your tissue samples, proteinase K will degrade it. Qiagen has a good animal tissue purification kit. The only thing I can recommend: incubate your samples at 65C, instead of 55C. Or you can leave the samples overnight at 55. Do not use more than 50 ug of tissue.
you need firstly to defrost the sample and then follow any protocol for genomic extraction from tissue
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