I have don't DNA isolation using CTAB method and got good precipitation in the isopropanol stage. But finally when I analyzed my samples in 0.8% agarose gel I could not find any resolved DNA fragment or any DNA in the well. Does the precipitation in the isopropanol stage confirm the presence of DNA in our sample? I have used RNAse treatment and hence I don't think it could be RNA. Eve if RNA was present why was it not shown up in the gel?
Thank you for answering this quiet basic question
That precipitation after isoprop is not for DNA or RNA it is protein precipitation, DNA and RNA is not visible as precipitate.You can check your sample concentration for DNA and protein(if possible then use Qubit) to get an idea about the quality of DNA, but if it is not seen in the gel then concentration is too low to do downstream assays.
Are you precipitating your nucleic acids with isopropanol??
Isopropanol/ethanol are used for precipating nucleic acids with different purposes such as purification of the nucleic acids. When you perform this precipitation, it's for sure that you'll lose quantity of your nucleic acids; however, in the final step to discard the supernatant (after centrifuge) you have to be really careful of not disturbing the pellet which was formed and contain your nucleic acids, otherwise you could lose totally your nucleic acids. You can quantify your nucleic acids (either DNA or RNA) using nanodrop system which is a really good and fast equipment to quantify your nucleic acids.
yes I did check with nanodrop and got it quantified. But I don't find the bands int he gel.
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