Dear Abdallh,

In general, the maximum plex number to achieve an acceptable amplification by rt-pcr is 5 plex. Although it theory 6 plex could be possible, maybe, y don't recommend you. You could combine amplicon size and fluorescence . I mean, amplify products with different size and use (for example 3 different fluorophores). (as an example “Rapid and Easy Identification of Capsular Serotypes of Streptococcus pneumoniae by Use of Fragment Analysis by Automated Fluorescence-Based Capillary Electrophoresis”). You could distinguish them by the combination of spectra and melting temperature. BUT, maybe it would be easier to perform two different reactions.

Dear Abdallh,

In general, the maximum plex number to achieve an acceptable amplification by rt-pcr is 5 plex. Although it theory 6 plex could be possible, maybe, y don't recommend you. You could combine amplicon size and fluorescence . I mean, amplify products with different size and use (for example 3 different fluorophores). (as an example “Rapid and Easy Identification of Capsular Serotypes of Streptococcus pneumoniae by Use of Fragment Analysis by Automated Fluorescence-Based Capillary Electrophoresis”). You could distinguish them by the combination of spectra and melting temperature. BUT, maybe it would be easier to perform two different reactions.

thanx a lot Hector D De Paz 

It is possible but could not be recommendable by low efficiency, is adequate doing it like Hector D De Paz said