I used DNeasy (Qiagen) and invitrogen DNA mini extraction kits with digestion modification, but I am not getting DNA's. So please anyone can give me the protocol or kit to extract DNA from 50 to 100 years old museum preserved/dried bird samples. Thanking you.
A phenol and chloroform extraction protocol is commonly used for older museum specimens.
It is possible that you are losing thea digestion with Proteina digestion with Proteinase K overnightase K overnight majority of DNA with these kits, even if you should still get an amount that could allow PCR experiments: you could try to warm at 37°C the elution buffer and use a smaller volume. otherwise you can try a digestion with Proteinase K overnight followed by a phenol-chloroform as Robert suggested. If samples are paraffin embedded however you shoud use a separate protocol!
Dear Robert Edward Wilson, I am using Footpad/toe pad samples, How much weight (mg) of sample I can take for further Phenol-Chloroform extraction. Have you got good DNA product.
How do you know you're not getting DNA? Are you quantifying your extracts or is it because you cannot get anything to amplify? You could have inhibition. I'd recommend soaking the toe pads in distilled water, overnight, and then extract using DTT and Proteinase K at 56C for 1-2 nights, and purify and concentrate using for example millipore columns.
If you are deciding whether there is DNA or not , only based on looking at gels after extraction, then you might be wrong. Many times you may not see anything on a gel but still have DNA enough to do a succesful PCR. On the other hand (at least with DNEasy kit), increasing incubation and elution times sometimes helps. Good luck !
Yes I've had great success extracting DNA from museum toe pads using Bioline Kits. I digest overnight at LEAST ensuring all the material is digested. If you are trying to visualise DNA on a gel you may not see it even if it is there. If you are trying PCRs say for mtDNA markers, you need to ensure you have very short fragment primers (150-200bp). Good Luck!
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