Archives: FAQs

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There is precipitate in my TwistAmp® Liquid E-mix. Can I still use it?

Some small amount of the phosphocreatine in E-mix can occasionally form a precipitate. This should not affect RPA reactions as there is a large excess of phosphocreatine in the E-mix. Vortex the tube to resuspend precipitate and spin down any remaining precipitate before using the supernatant.

I’ve tested some primers with one type of TwistAmp® kit; will they work with another type of TwistAmp® kit?

Maybe. If you have developed primers using a TwistAmp® Basic kit, you will also need to include a probe if you are going to use a TwistAmp® exo or nfo kit. This and other factors mean that an optimal primer combination for a gel-based approach vs a fluorescence or lateral flow approach may not be the same. In general we …

I’ve tested some primers with one type of TwistAmp® kit; will they work with another type of TwistAmp® kit? Read More

I want to run multiple experiments using TwistAmp® Liquid reactions; can I make a master mix and store it?

Oligos, 2x Reaction Buffer, water, dNTPs and 10x E-mix can be combined and stored on ice for a minimum of hours, and over-night in many cases (any effects on performance will be assay dependent and should be assessed independently by users). Once the 20x Core Reaction Mix is added short term storage on ice (assay dependent again) should also be …

I want to run multiple experiments using TwistAmp® Liquid reactions; can I make a master mix and store it? Read More

I keep getting false positive results, what is the likely cause of this?

If positive results are seen in no template controls it is commonly caused by contamination, particularly, if the positive control oligonucleotides provided in the kit display false positives when no template is added. Contamination can occur if you are pipetting high copies (10^6 /µl or above) of template in the same area as carrying out RPA testing. We strongly recommend …

I keep getting false positive results, what is the likely cause of this? Read More

How much of my probe do I have to use?

The recommended concentration of probe in the reaction is 120nM. However, some TwistAmp® reactions benefit from being used with slightly different amounts of probe. Testing different concentrations of probe (from 50nM to 150nM) will help to optimise the performance of probe based detection formats of a given assay.

How much of my primers do I have to use?

The recommended concentration of primers in the TwistAmp® Basic reaction is 480nM each. In the TwistAmp® exo, TwistAmp® fpg, and Twist Amp® nfo kits the recommended concentration of primer is 420nM each. However, the performance of some primer pairs can be improved by slightly varying their amounts in the reaction and a titration strategy (from 200nM to 600nM each) can …

How much of my primers do I have to use? Read More

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