I've used a SpeedVac for concentrating pDNA after extraction with a commercial kit and lost a lot of material. While the volume was reduced to around 20%, the increase in concentration was very low, less than 10%.
Can someone explain this and/or give hints on how to avoid that?
I know that ethanol precipitation is an option, but I would like to avoid that, because I'm going to use this for electroporation and don't want to risk having any residual salts.
the usual reason is that the sample is not frozen so it boils and splatters material up the sides of the tube. Make sure the sample is solid and remains solid during spinning and that the tubes are spinning before turning on the vacuum
you also must be careful that when you concentrate dna the salts in the buffer get concentrated as well and then you have to make sure that the zero sample blank on the spectrometer you are using to measure the dna concentration is appropriate to the sample that you are measuring or the concentrations measured will be wrong
I have also experienced that, only with small amounts of the DNA and only when I used QIAamp (QIAGEN). Ultimately I decided to omit the step. Nevertheless, you can possibly try to elute DNA in a different buffer or just in water and reduce the temperature during the concentration.
What are you using to measure the DNA concentration?
What is the input sample material?
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