Hi all,

I worked with RAD libraries with Bryophyta and we successfully used SBFI restriction enzyme. We used same protocol (because we have all adaptors) to vascular plants (Carex, Pedicularis) but without success - it seems that restriction is not working well. Is here somebody who uses successfully sbfI to plant genomes? Which ratio enzyme/DNA amount you use?

And the second question, mosses have small genomes around 2C 1pg, we pooled 58 samples without any problem. We used Hiseq Illumina and got approx. 2 mil reads per individual and 78 thous. rad loci, mean coverage 60-100 per individual. Pedicularis has 4pg, but probably the fewer restriction sites, how to pool it to get similar results? In only paper which I found, they pooled 24 Pedicularis samples for RAD, but they use PstI, what can be different.

Thank for your suggestions

Eva