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RPA Reaction Buffer (2X) - FAQs

RPA Reaction Buffer (2X) - FAQs

View additional product information for Lyo-ready Enzymes and Reagents for Recombinase Polymerase Amplification (RPA) - FAQs (A72126, A72125, A72128, A72127, A72124, A72123)

6 product FAQs found

Can I use PCR primers for target amplification by the Lyo-ready RPA Kit (Cat. No. A72127, A72128)?

PCR primers can be used with the Lyo-ready RPA Kit if they are at least 30 nt long. RPA primers can be designed using Primer3.

Find additional tips, troubleshooting help, and resources within our PCR and cDNA Synthesis Support Center.

Can I lyophilize reagents from the Lyo-ready RPA Kit?

All RPA reagents (including Lyo-ready T4 UvsX Protein (Cat. No. A72124), Lyo-ready T4 UvsY Protein (Cat. No. A72125), Lyo-ready T4 Gene 32 Protein (Cat. No. A72123), Lyo-ready Bst DNA Polymerase (Cat. No. A56655), and RPA Reaction Buffer (2X) (Cat. No. A72126)) are lyo-ready, ensuring that enzymes and buffer formulations are glycerol-free. However, the formulations do not contain excipients needed for the lyophilization process.

Find additional tips, troubleshooting help, and resources within our PCR and cDNA Synthesis Support Center.

I noticed that my Lyo-ready RPA Kit is frozen after storage at -20 degrees C. Is it still good to use?

Lyo-ready RPA Kit and stand-alone reagents come in a storage buffer that does not contain glycerol. Product freezing doesn't affect its performance or stability. We do not recommend freeze-thawing Lyo-ready RPA Kit and stand-alone reagents for more than 20 cycles.

Find additional tips, troubleshooting help, and resources within our PCR and cDNA Synthesis Support Center.

What is the temperature limit to perform an RPA reaction using a Lyo-ready RPA Kit (Cat. No. A72127, A72128)?

Lyo-ready RPA Kit can amplify DNA in temperatures ranging from 37 to 45 degrees C. To ensure the best performance, we recommend using the optimal reaction temperature of 42 degrees C.

Find additional tips, troubleshooting help, and resources within our PCR and cDNA Synthesis Support Center.

Can I use my reverse transcriptase with a Lyo-ready RPA Kit (Cat. No. A72127, A72128) for RT-RPA?

We recommend using SuperScript IV Reverse Transcriptase (Cat. No. 18090010, 18090050, 18090200) for the best results in RT-RPA. Guidelines for use of SuperScript IV Reverse Transcriptase in RT-RPA application can be found in the Lyo-ready RPA Kit user guide. Other reverse transcriptases such as Maxima H Minus Reverse Transcriptase (Cat. No. EP0751) have also been tested and can be used in RT-RPA application. The list of Lyo-ready reverse transcriptases can be found here: thermofisher.com/lyo-ready.

Find additional tips, troubleshooting help, and resources within our PCR and cDNA Synthesis Support Center.

Can the Recombinase Polymerase Amplification (RPA) reaction volume be scaled up for Lyo-ready RPA reagents?

Yes, the reaction volume can be scaled up to 50 µL from the recommended 20 µL. Scaling up the reaction volume more than 4-fold may affect performance.

Find additional tips, troubleshooting help, and resources within our PCR and cDNA Synthesis Support Center.