I used 0.7% Agarose gel for gel run A, 1% for B and 0.8% for C. Gel run A had samples processed (DNA extracted) by grinding tissue with liquid nitrogen and mortar pestel, run B and run C with homogenizer.
DNA is a long thin molecule so any isolation method using grinders or homogenisers will lead to broken up DNA and after any dry grinding it is a good idea to use the first liquid added to the crude extract containing EDTA in order to inactivate nucleases during the purification process. If your next step is pcr then degradation of the dna which is larger than your pcr product then I would expect your pcrs to work well
Paul Rutland we're hoping to send the DNA samples for Bisulfite-based methylation analysis. Would fragmented DNA be an issue then?
Hasib Ahmed I do not know the requirements for bisulphite analysis. It might be a good idea to contact tech support at the company doing your analysis and check with them what the qulity criteria are.
Hi, Hasib Ahmed, your photos show a smear that means your DNA is degratated. May be it is a bad conservation of the tissu or the way how you take organs. Normal DNA has a high molecular weight. On 08% agarose gel you can see a band on the top of your DNA ladder
For me the best way to get DNA, is to use proteinase K. Here the article reference : Simplified mammalian DNA isolation procedure
Nucleic Acids Research, vol 19, no 15 4293 1991
@Guy Longepied we collect placental tissue samples from hospitals in plastic vials containing RNAlater solution, and store them at -20. We use the Qiagen Tissue kit with supplied Proteinase K to extract the DNA.
It seem that you want to work with RNA and DNA. The RNA later is use to keep tissu for RNA extraction.
When you collecte placental tissue, divided it in 2 small parts : one for RNA extration and one for DNA extraction. The important thing is to freeze immediately in liquid nitrogen to avoid any degradation with DNAse or RNase
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