I'm currently working on a cloning protocol and amplified my inserts through PCR and purified by gel extraction.
After agarose gel extraction, I got a 60 uL DNA sample way too diluted for downstream applications (LIC reaction) and decided to concentrate the sample in a vacuum concentrator.
It happens that i accidentally dried out the sample (the process was carried out at 62°C). I then tried to add 40 uL of miliQ water to the dried out tube. Nanodrop quantitation showed I had enough DNA for my application but now I'm worried: could the DNA be too damaged and unreliable as a cloning insert? The 260/280 ratios are the same now as they were after the gel extraction (i.e. before I dried it out). Does this indicates the sample can still be used as a cloning insert?
dna is very stable and I would not expect any problems. There are many millions of molecules and most of them will be undamaged and will work well. You accept a degree of taq damaged amplimers in pcr and you will probably not notice the difference with just a few more damaged amplimers
dna is very stable and I would not expect any problems. There are many millions of molecules and most of them will be undamaged and will work well. You accept a degree of taq damaged amplimers in pcr and you will probably not notice the difference with just a few more damaged amplimers
As it was said before, I think you can use your PCR products, DNA is very stable.
There should be no problems. Sometimes overdried DNA shows solubility issues and, in that case you may want to try some diluted NaOH to increase solubility.
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