Hi everyone, I am trying to run the agarose gel to identify my RNA integrity. One rna sample concentration is around 37ng/ul, the other one from liver tissue, the [C] is around 600ng/ul.I am wondering how much loading dye (6X) and how much RNA sample would you suggest for me to load?
I been looking around trying to find answer, but seems like there's no stander, it's a little confusion for a beginner like me. Before, my RNA sample was around 30-60ng/ul, I just took 7.5ul rna mixed w/ 2.5ul 6X dye, they came out fine, do show some smear.
Anyway, this time, I want to run those two samples together. so one more q, do i have to convert the concentration to the same i mean around?
Really appreciate
In gel you only see the integrity of your RNA , therefore no need to equalize the concentration to same to all samples. 2uL RNA and 2uL Dye will be good to check the integrity. Few more thing I would like to add dissolve your RNA in a RNA storage solution from Thermo Scientific and also use RNA loading dye from same vendors.
Agarose gel is basically run to verify the quality of RNA. Equalize the concentration of both the samples. Add loading dye such that final concentration of loading dye is 1X. Equalization of the samples helps to compare the intensity of RNA bands among different samples. Generally I use 500ng of RNA sample.
Hi
the most important thing to keep in mind is the use of denaturing loading dye. If not your RNA will migrate like the degraded one. If you are just interested by knowing the quality of your RNA you do not need to equalize the quantities.
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