Hi.
I'm preparing RNA-seq libraries and I measure the concentration using Qubit assay. I got good values (around 6-8), but when I run the gel, nothing appears, only the ladder. Does it mean I don't have anything in the sample? And how can I get good concentration values? Thank you very much.
Hi. Thanks for your answer. 6-8 ng/uL. I load 4 uL of my libraries with 2 uL of loading buffer along with 1 ng of the ladder, in a gel with SybrSafe, and then visualize them with UV. I usually see the libraries on a gel (as most people do) to check the quality and fragment sizes.
Miquel!! Era una pregunta general.. No solo para ti..jeje .. Todo bien?
Hola José, no sabia que ahora estas en USA, volviendo a tu pregunta, será que tienes un problema puntual de contaminacion con RNasas? Ves algún manchon (smear) concentrado al final del gel como si tuvieses un monton de micro RNA?
Si.. ya ves. Por aqui ando. La verdad es que no aparece nada en el gel. No se por que el Qubit me da buenos valores de concentracion. Un saludo!
Sorry to say but communicating in a language not understood by many in researchgate community is kind of impolite. The whole purpose of joining this community is to learn from each other's experience and knowledge and that purpose is not served if common language is not used for communication.
Sorry about that. They are colleges from my previous center and it is difficult for us to communicate in other language. Thanks Nina for your suggestions. I think Qubit reagents are not contaminated since I use the standards too. Most likely the library may be degraded as I can't see it. Thank you.
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