I have a problem with my DNA ladder .. I found that the separation ladder sizes of my DNA ladder are poor. the agarose gel I use is 1% with TBE buffer. I have tried a few times with lower voltage 50Volt to run my electrophoresis gel but still, the separation is not separated well. Is there any suggestion on how to improve my DNA ladder? thank you
Dear Siti, just increase the concentration of the gel. The separation of DNA ladder depends on the percentage of the agarose gel. If you have a PCR product in the of range 100bp and more you can use the 1,5-2% gel.
If you want to resolve the DNA with size below 100 bp you can try 2-4% of agarose in 1X TAE.
Another better alternative is to resolve the sample in 8% native vertical polyacrylamide gel in 1X TBE.
Your problem is not voltage but most likely you do not let it run long enough. It seems like your migration front only goes to the middle of the gel.
Let it run until it reaches 1cm from the bottom of the gel.
TBE should be fine, but you can go for 1.5% or 2% agarose for better resolution as someone else mentionned.
Thank you all for the your suggestions. Will take notes and try it out
I have seen something like that when running gels at too high voltage.
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