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DNA marker not separating very well

DNA marker not separating very well - Agarose Gel Electrophoresis Forum

DNA marker not separating very well - Agarose Gel Electrophoresis Forum. A forum to discuss electrophoresis staining, troubleshooting, and development of films for DNA and RNA gels.


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Old 08-07-2011, 01:00 PM
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Default DNA marker not separating very well



I have been having with my DNA marker...It does not separate as in the diagram gave to me by the supplier...it is a 1kb DNA marker, electrophoresis is done with 2% agarose get with 1x TBE buffer with 70volt for an hour. The band is very thick and look like it does not separate. Help needed.
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Old 08-09-2011, 03:50 AM
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Default Re: DNA marker not separating very well

Try running it in a less concentrated gel (for instance 1%) and for a bit more of time. Raising the voltage a little wouldn't harm. If it still doesn't separate then call the supplier for troubleshooting.

Good luck.
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Old 08-09-2011, 12:45 PM
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Default Re: DNA marker not separating very well

We've experienced that if you dilute the ladder a bit before you use it, the bands will separate better/or you'll be able to see the bands better.
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Old 08-10-2011, 12:06 AM
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Default Re: DNA marker not separating very well

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Originally Posted by luisillo View Post
Try running it in a less concentrated gel (for instance 1%) and for a bit more of time. Raising the voltage a little wouldn't harm. If it still doesn't separate then call the supplier for troubleshooting.

Good luck.
I have contacted the supplier, they said the dna ladder i used contain Orange Dye which is not compatible with the GelRed I use to stain the agarose.

but I will try and run it again with lesser agarose concentration and see if there are any changes.

Thanks for replying.
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Old 08-10-2011, 12:08 AM
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Default Re: DNA marker not separating very well

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Originally Posted by Aurora Borealis View Post
We've experienced that if you dilute the ladder a bit before you use it, the bands will separate better/or you'll be able to see the bands better.
hello,

it stated that the DNA ladder is ready to use with premix orange G loading dye, can I still dilute it?...if so...can teach me how?..

Thanks for the information .
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Old 01-15-2013, 10:52 AM
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