FAQ: Do I need to change my promoter sequence to use CleanCap® Reagent AG?

The T7 RNA promoter sequence itself (5′ TAATACGACTCACTATA 3′) does not need to be changed as the kit still utilizes T7 RNA Polymerase.  However, the two initiating bases (+1 and +2) that are just downstream of the T7 promoter need to be “AG”.  (The +1 base following the standard T7 RNA Polymerase Promoter is always a “G”.  However, the +2 base may be any base (N)). 

Original promoter with initiating base +1 (G) and +2 (N)
5′ TAATACGACTCACTATAGN 3′

New sequence required for use with CleanCap Reagent AG (with initiating bases AG)
5′ TAATACGACTCACTATAAG 3′