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-   -   DNA marker not separating very well (http://www.molecularstation.com/forum/agarose-gel-electrophoresis-forum/83312-dna-marker-not-separating-very-well.html)

myfaridah 08-07-2011 01:00 PM

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.

luisillo 08-09-2011 03:50 AM

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.

Aurora Borealis 08-09-2011 12:45 PM

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.

myfaridah 08-10-2011 12:06 AM

Re: DNA marker not separating very well
 
Quote:

Originally Posted by luisillo (Post 435600)
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.

myfaridah 08-10-2011 12:08 AM

Re: DNA marker not separating very well
 
Quote:

Originally Posted by Aurora Borealis (Post 435603)
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 :).

carteyroabl 01-15-2013 10:52 AM

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