FAQs

Yes, if you have used TwistAmp® Basic or nfo reactions, you can store your amplicons at 4°C for short periods of time and -20°C for longer periods of time. No, if you have used TwistAmp® exo (RT) reactions. The exonuclease III is likely to digest the amplicon if the reaction is not processed rapidly to inactivate any such activity.

The Twista® is not Windows 8 compatible, so you will have to install an emulator of an earlier version of Windows (XP, Vista or 7) in order to get it to run.

Yes. The measurement of change of fluorescence can also be used as an endpoint read-out. In this case the fluorescence at the start and at the end of the reaction is compared: an increase of fluorescence signifies a successful amplification event.

Yes. It is possible to perform more than one amplification reaction simultaneously in the same tube. However, not all primer pair combinations will work equally well with each other in multiplexing and this format therefore requires careful primer design. Note that the total amount (nmols) of oligonucleotide in the reaction should not significantly exceed that stated in the protocol; if

Yes. If you wish to set up multiple reactions, you can make a master mix. If you are screening different DNAs, the resuspension buffer, primers and probe if used can all be mixed together and added to freeze dried reactions to resuspend them. Different DNAs can then be added to reactions before they are started with MgAc as usual. If

Yes. You can use two modified primers for lateral flow if you wish, but we recommend using a probe and a TwistAmp® nfo kit. This is because, as with PCR, RPA is subject to primer-dimer formation, so you can get primers cross reacting and giving false positives if you do not have perfectly designed primers. TwistAmp® nfo probes avoid this

Yes, if you are using TwistAmp® Basic, nfo or fpg reactions. However the reaction components are activated as soon as magnesium is added. By pipetting magnesium acetate into the reaction last, which we recommend, for example by adding it to the lids of strips and spinning it into the reactions, you ensure that reactions start simultaneously and you minimise the

Small changes in the sequence of a primer can improve RPA performance. Any given primer can be optimised by slightly varying its length (by single nucleotides) and position (keeping the length but shifting their location in 1bp increments) and re-testing its activity.

Yes, unless you are using the TwistAmp® exo kit or exo RT kit. The exonuclease present in the reaction mixture will digest most of the amplification product once amplification has ceased. For the analysis of amplicon, use the TwistAmp® Basic kit. It is also possible to analyse products on gels generated from the TwistAmp®nfo and TwistAmp® fpg kits. NB The

No. Adding one primer or probe before another will bias the formation of recombination filaments towards whichever oligonucleotide is added first. This is why primers and probes are added simultaneously.