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quiaquick gel extraction troubleshooting


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#1 labgirl14

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Posted 12 January 2015 - 10:52 AM

Hi, I am new to the lab this year and just did my first gel extraction last week. I made a couple of mistakes and was wondering whether or not these have significantly damaged my end products and would need to redo yhe gel extraction? I would run a gel to check but i just wanted to clarify. The mistakes were:
1. Melted agarose at 100degrees celcius not 50degrees
2. Centrifuged after adding isopropanol but then mixed with pipette after centrifuging before tranfsferring to minicolumn.

Thank you in advance

#2 CPRES

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Posted 12 January 2015 - 11:09 AM

As you mentined, you should run a minigel to check the integrity and amount (it may not even be there) of your DNA. You can check the quality of DNA by spectrophotometry. And if it is there and looks good, as long as you are not using it for some sophisticated or finicky technique (i.e. mouse embryo microinjection), you should be ok.

 

However, over the years, the most important rule of mol bio techniques I have learned is mistakes are cumulative. You or someone else will use your reagents when you don't even remember when and how it was produced, and may run into unforeseeable troubles. So it is best to nip the problem in the bud and repeat a nice and clean procedure. You will be satisfied, and sleep well all year, I guarantee it!


So. Now that you have your first ever question on bioforum answered (or not), mail yourself your username and password so you don't forget them, and then come back soon to update us on how it all worked out. That's how you build Karma in science.





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