The viruses do drop RNA segments and when they do, they are
noninfectious. If you have a high proportion of viruses lacking a
segment or two in any given viral stock, and then run a gel like this
one, you will see sub-molar ratios of specific RNA segments. Never
missing a segment – such a virus could not propagate on its own, e.g.
without a helper virus.

This gel system worked well, but it was finicky. Everything had to be
done just so, or else nothing. I had many smeared gels that never saw
the light of day.

The viruses do drop RNA segments and when they do, they are
noninfectious. If you have a high proportion of viruses lacking a
segment or two in any given viral stock, and then run a gel like this
one, you will see sub-molar ratios of specific RNA segments. Never
missing a segment – such a virus could not propagate on its own, e.g.
without a helper virus.

This gel system worked well, but it was finicky. Everything had to be
done just so, or else nothing. I had many smeared gels that never saw
the light of day.

First, I only see 7 bands in the MD gel. And that 2 band in R3 looks a lot like L-3.

Not knowing anything about the virology and just looking at the gels, one might argue that R3-1 => L-2. R3-2 => L-3. And R3-3 => M-3, which would mean that R3 is lacking the 1 band and has 2 copies of the 3 band. Does the virology support this type of Ho? Do scrambled cultures drop segments out and/or double up on other segments?

Alternatively, if what we are seeing is M-1 and M-2 smeared, one could conclude the R3-1 and R3-2 are M-1 and M-2 properly separated (i.e. artifact). Is that the fast end of the gels at the top? Don’t you expect more wobble there?

As to bands 4/5, one interpretation is that R3-4 is common to all three gels and L-“4″ is really M-5, which in the Lee strain moves faster than the M-4 IOW, the 4/5 doublet could be flip-flopped in the Lee/Md strains. Do such doublets ever flip flop in these viral RNAs?

BTW, as these comments probably suggest, I am really jealous. At about the same time you were getting these beautiful results I was running gels (DNA) and getting spotted film that looked like shots from an X-ray telescope of galaxies in the deep universe! I knew I shoulda' gone into astronomy.