Basic Information

RPA

1) Primers must be 30-35 bases

2) Works best at constant temperature (37-39°C)

3) Amplicons of 80-400bp are preferred

4) TwistAmp® exo Probe required - see overleaf

Set-up (single-plex)1

1) Prepare reaction mix in 1.5ml tube:

Primer A (10μM)    2.1 μl

Primer B (10μM)    2.1 μl

TwistAmp® exo Probe (10μM)    0.6 μl

Rehydration Buffer   29.5 μl

Template and dH2O   13.2 μl

(Total Volume   47.5 μl)

Vortex and spin briefly

2) Add reaction mix to freeze-dried reaction. Pipette to mix.

3) Add 2.5 μl of 280mM MgAc (supplied) and mix well to start reaction.

4) Place strip in Twista® and start run: 37-39°C, 20 minutes.

4b) For low template copy number, remove strip after 4 minutes, vortex &

spin briefly, replace in Twista®.

WARNING: RPA REACTIONS START AT ROOM TEMPERATURE AS SOON AS