DNA marker not separating very well

[myfaridah]

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]

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]

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]

Originally Posted by luisillo

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]

Originally Posted by Aurora Borealis

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]

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