Mixing RT-PCT master mixes and plates - Best way to do it? (Nov/27/2008 )

For a one-step RT-PCR reaction how would be best to mix the reagants in the master mix tube? Vortexing or pipetting... followed by a spin-down?

Also I've previously done two-step RT-PCR using pre-made cDNA on the plate. We used to seal the cover down, vortex the plate then spin it down. Would this method work okay for a total RNA starting template? Or alternatively can suffucient mixing of the MM and template be obtained during pipetting out the MM?

I'm interested to see people's methods and reason for choosing them.

Thanks

For a one-step RT-PCR reaction how would be best to mix the reagants in the master mix tube? Vortexing or pipetting... followed by a spin-down?

Also I've previously done two-step RT-PCR using pre-made cDNA on the plate. We used to seal the cover down, vortex the plate then spin it down. Would this method work okay for a total RNA starting template? Or alternatively can suffucient mixing of the MM and template be obtained during pipetting out the MM?

I'm interested to see people's methods and reason for choosing them.

Thanks