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Topic: Need help rehydrating DNA sample for PCR/gel electrophoresis  (Read 2727 times)

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Offline DNAissues

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Need help rehydrating DNA sample for PCR/gel electrophoresis
« on: April 03, 2016, 12:19:10 AM »
Hi!

I am doing an experiment for my Diploma Project where I analyze and compare a few samples of DNA (all from the same source, my saliva) that have stayed outside for various amounts of time before being frozen.

I chose to let the samples stay with no lid on but I am now facing the problem of evaporation, and I have no clue how to proceed in rehydrating the pellets after they have been thawed.

I have tried researching on this but found no credible/relevant sources. I am not very experienced in this subject so I might have been searching for the wrong things so if anyone could direct me to some protocol or help in any other way it would be extremely appreciated!

If there is anything I would need to clarify, please ask. I have been using a standard protocol with Chelex.

I apologize if this is not the right forum to post in, but it seemed the most relevant.

Thank you in advance!

Offline Arkcon

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Re: Need help rehydrating DNA sample for PCR/gel electrophoresis
« Reply #1 on: April 03, 2016, 05:24:03 AM »
So you have a dried DNA pellet, and its hard to rehydrate?  Why not start with whatever standard you're going to run, dry it the same way, and see if it rehydrates adequately in the PCR/PAGE buffer?
Hey, I'm not judging.  I just like to shoot straight.  I'm a man of science.

Offline Yggdrasil

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Re: Need help rehydrating DNA sample for PCR/gel electrophoresis
« Reply #2 on: April 03, 2016, 04:07:35 PM »
You should be able to just add some aqueous buffer (e.g. TE buffer) and it will dissolve.  Depending on the size of the DNA and the amount, it may require some time for the dissolution to take place, however.  Try adding the buffer, mixing, letting it sit at room temp or 37oC for 15 min, come back and mix again, and repeat until you see the pellet dissolve (depending on whether you had any other contaminant like protein, you may not see the pellet fully redissolve).

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