Sequencing RNA

[barrmur]

Hi all,

Just wondering if anyone has any experience of the following. I want to sequence a single fragment of RNA to confirm it is what I think it is. Any suggestions on how to go about it. The major problme is it is only 25 bases long. I can convert to DNA and have a 7bp primer but not sure where to go from here. I have access to illumina, pyromark Q24, ABI 3730. I have tried cloning the fragment with no success. Any other suggestions.

[malachig]

Have you considered making a concatamer and then cloning that? Analogous to sequencing of SAGE libraries by Sanger sequencing.

[barrmur]

Nice idea, might give that a go. Cheers.