Bioanalyzer problem-rna ladder 600 nano

[Arianna Cerana]

Hello everybody!
I have a question for you.
Do you ever seen a ladder (RNA 6000 nano) like this? What can be the causes?
I have prepared a fresh gel-dye mix and the electrodes of the instrument have been cleaned last week, no other analysis was made in this period.
The gel matrix, the RNA dye concentrate solution and the ladder are expired two months ago, it can be this the problem?

Thanks in advance

[Ben3]

Hello Arianna Cerana

I've used expired reagents before without any issues, but it still could technically be the issue. Most of the issues I've had were from not heat denaturing properly or issues during the dye mix.

[Arianna Cerana]

Hello Ben3, thanks for your answer!!
When you say "not denaturing properly", you mean the ladder right?
Maybe I can denature it another time?

Thank you so much for the help

[Ben3]

Arianna Cerana. Yes, I've had issues with denaturing the ladder in the past. I think it may have been the thermocycler I used. But most of my issues were with mixing the dye. They may not be the issues, but I figured I would share in case it was. Best of luck!

[Arianna Cerana]

Thank you so much!!
I can prepare a new gel-dye mix and see what happens.
Thanks again for the support and the help!!

[Arianna Cerana]

Thank you so much celinedion!! I re-prepared the gel-dye mix and it works!!
Maybe also the samples can influence the conditions? Because I had problems only with a batch of samples.