() The components of the pGRFP plasmid vector

Copyright information:Taken from "A dual-fluorescence reporter system for high-throughput clone characterization and selection by cell sorting"Nucleic Acids Research 2005;33(5):e49-e49.Published online 14 Mar 2005PMCID:PMC1065264.© The Author 2005. Published by Oxford University Press. All rights reserved The main components are the pUC origin of replication, ampicillin resistance marker and a fused GFP-DsRed gene separated by a linker. The linker region is shown with six amino acid linkers (SGSGSG and GSGSGS) on either side, M13 forward and reverse priming sites, and EcoRV, NotI and SalI sites. () Flow-cytometry configuration for dual-fluorescence quantification and sorting. A 488 nm laser excites the fluorescent proteins in individual suspended in the flow stream. The flow cytometer is configured to trigger either on forward scatter or GFP fluorescence. The fluorescence is split using a 550 nm dichroic long pass beam splitter. The green fluorescence is filtered through a 560 nm short pass filter before detection. The red fluorescence passes through a 590 nm long pass filter before detection.