Dear all, I would like to thank you for your quick and helpful replies concerning AFLPs. As many of you were interested in the answers, here comes a short summary of them: Purification of selective PCR products: -most people do not do a sephadex purification because one might loose fragments -those who do a sephadex purification report that this leads to a reduction of primer-dimers and a better resolution and clarity in the range from 75-500 bp -one can also try a purification of PCR products following precipitation protocols for sequencing Sample volume: -1-3 [cid:image001.png@01C898A1.8EAF7840] multiplexed PCR product in 10-15 [cid:image002.png@01C898A1.8EAF7840] l total volume (HiDi and Standard) Injection time: -10-40 (or even 90) seconds PCR protocol: -use of other/better Taqs or PCR kits (see also Trybush et al, Can. J. Bot. 84: 1347-1354 (2006)). -20-30x dilution of the multiplexed selective PCR product will lead to better/higher peaks Again, many thanks to all of you who helped, and good luck with your AFLPs! Simone Steffen Institut für Spezielle Botanik und Botanischer Garten Bentzelweg 9a 55099 Mainz Germany phone: +49 6131 3922624 fax: +49 6131 3923524 mail: ssteffen@uni-mainz.de ssteffen@uni-mainz.de