RNA ligation - (Dec/03/2001 ) I'm not sure if you can do single
stranded RNA ligation, but I have another idea. What if you did
first strand synthesis on the RNA. Then, do a second strand
synthesis (protocol in maniatis or current protocols). Next,
blunt the ends of the double stranded cDNA you produced (just to
be sure). Next, make double stranded linker DNA that has a
specific primer site in it and do a ligation with your double
stranded DNA. Finally, do a PCR reaction with this ligation as a
template and the primer you designed into the linker DNA, and a
specific primer you choose from sequence you already know. You
will probably get several bands, but hopefully you can estimate
the size of the band you want. Gel purify that band and sequence
it directly or subclone it. There are protocols in maniatis and
current protocols on how to do each of these steps. In theory, I
think this should work- any thoughts from anyone else? Is there
such thing as RNA ligation? I have a RNA virus genome to
sequence but being