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Dissolved Primer pellet in pure ethanol instead of water


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#1 Moebius

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Posted 09 March 2011 - 06:54 AM

I accidentally grabbed the ethanol bottle instead of the water one. The ethanol bottle has 100% ethanol. I noticed the stock solution didn't freeze at -20 C and I didn't get any results for my PCR reaction. I was wondering if I can just pipette out the ethanol and add the appropriate amount of water into the primer tubes. I can still see the pellet, but some of the pellet has gotten bigger in area. I really appreciate any help in advance! Thank you!

#2 perneseblue

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Posted 09 March 2011 - 08:16 PM

I would first spin down the tube to make sure all the primer has settled.
Only then, I would carefully pipette out the ethanol. To remove any remaining ethanol, I would leave the tube on the bench to dry.

If you have a vacuum desiccator, I would pop the tube in and have the ethanol evaporate away. That is much safer than attempting to remove the ethanol by a pipette.

A 50 C incubator could also be used to evaporate the ethanol.
May your PCR products be long, your protocols short and your boss on holiday

#3 Moebius

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Posted 10 March 2011 - 08:40 AM

I would first spin down the tube to make sure all the primer has settled.
Only then, I would carefully pipette out the ethanol. To remove any remaining ethanol, I would leave the tube on the bench to dry.

If you have a vacuum desiccator, I would pop the tube in and have the ethanol evaporate away. That is much safer than attempting to remove the ethanol by a pipette.

A 50 C incubator could also be used to evaporate the ethanol.


Thank you! I got positives for almost all the primers when I ran the reaction last night. :) I will definitely use the water bath in the future. I was wondering if its too dry, would it damage to primers if I leave it in the water bath for too long? I know DNA doesn't denature at 50 C, but I just want to be sure. Thank you and I love your avatar! I am still trying to figure out why the last two primers didn't work.




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