Zero Blunt™ TOPO™ PCR Cloning Kit, with One Shot™ MAX Efficiency DH5α-T1^R E. coli, 50 Reactions - FAQs

Can TOPO TA cloning vectors and Zero Blunt cloning vectors be used for in vitro transcription to generate probes? Can they be used to do in vitro translation?

Yes, the TOPO TA cloning and Zero Blunt cloning vectors can be used for in vitro transcription. The T7, SP6, and T3 promoters contained in these vectors are active phage promoters, and the corresponding polymerases will transcribe RNA from these promoters.
These vectors are not generally recommended for in vitro translation because most of these vectors contain one or more translational initiation signals (ATG) downstream (3’) of the promoters contained in them, within the multiple cloning sites, which may interfere with attempts to translate an open reading frame in the insert.