In DNA isolation, by mistake i used 100% ethanol wash in place of 70%. DNA band in agarose gel appeared fine enough.
Is there any chance of getting low quality sequencing result of those samples?
Unfortunately, that could results in poor sequence reads. The 70% ethanol is using for get rid of excess salt during DNA extraction, so you may provide well-prepared template for sequencing.
It might denature your sample if stored for long duration and may effect the sequencing results as well.
If the DNA is still precipitated, just do a wash with 70% ethanol and proceed as usual. If you already dissolved the DNA, you could precipitate it again, then wash the pellet with 70% ethanol, dry it and take it up in low salt buffer.
I disagree with Jyotsna: DNA is not desaturated by salt; however, as pointed out by Sepher Abdolahi, excess salt will hamper the sequencing reaction
Thanks for your valuable comment and I have already dissolve the DNA in Elution buffer and using this sample PCR gave good result (checked by agarose gel electrophoresis).so their is any chance of getting low quality sequencing result of those PCR product? I am attaching the gel image of those sample.
I guess there will not be any issue hereafter since you got product in PCR now.
It's not ideal to use 100% instead of 70% as you won't get excess salt removal. As others mentioned, you can re-wash your DNA (you will lose a bit of yield as you will need to re-precipitate). It may still work for sequencing, it will depend in part on how salty your samples were to start with, and if/how much you are diluting them for sequencing. Good luck!
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