Hello everyone
I want to know some detailed method how to run my total RNA to check its integrity by running 1% agarose gel.
Our lab has only regular agarose gel, sybr green with loading dye.
If I tried like below
2ul of total RNA +8 ul of water + 2ul of loading dye with sybergreen= total 12ul
Run the 12ul on the 1% regular agarose gel at 100Voltage with fresh buffer.
But I was not able to see sharp 2 bands for 28S an 18S.
There were some faint and little smeared 2 bands for 28S and 18S.
I am not sure that's because my RNA was degraded before or during running on the gel with high voltage.
Has anyone have a tip for running RNA on the regular agarose gel?
Hello
You should be able to see sharp bands with a standard agarose electrophoresis. To estimate the integrity of your samples based on RIN value, you may use a PDF file I sent you as an example.
Hi Hyunsook Moon ,
Check this thread, it'll help:
https://www.researchgate.net/post/Can_anyone_help_me_with_gel_electrophoresis_of_total_RNA
Cheers!
Thread Alejandro Martin has shared is very useful, but instead of 1% try to increase agarose concentration up to 2%. In my case, I found that 1.5% of Agarose in 1XTBE gives the best resolution.
Good luck!
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