PCR-genotyping of barley seedlings using DNA samples from tissue prints

Screening of large numbers of plant samples with polymerase chain reaction (PCR)-based techniques is a central task in marker-assisted plant breeding and molecular genetics. While PCR and electrophoresis have been streamlined as a result of high-throughput analysis methods, the collection of plant material and its processing to make it accessible for reliable amplification reactions remain labour-intensive, costly and not amenable to automation. As an alternative to traditional extraction protocols, DNA can be bound to specially treated paper (FTA-paper). For PCR analysis, instead of being added as a liquid suspension, DNA is then released from small paper disks that are immersed into the reaction mixture. In the present study, several parameters were investigated for the successful amplification of various single-copy sequence characterized amplified region markers from barley tissue prints. The results show that the FTA-paper approach offers a quick, reliable and low-cost alternative for PCR screening even in a crop plant with a large genome.