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  6. Invitrogen™ Lyo-ready Enzymes and Reagents for Recombinase Polymerase Amplification (RPA)

Invitrogen™ Lyo-ready Enzymes and Reagents for Recombinase Polymerase Amplification (RPA)

Catalog No. A72127
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Quantity:
100 Reactions
120 μg
1600 μg
2000 μL
Product Type:
Lyo-ready RPA Kit
Lyo-ready T4 Gene 32 Protein
Lyo-ready T4 UvsX Protein
Lyo-ready T4 UvsY Protein
Reaction Buffer
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Catalog No. Quantity Product Type
A72127 100 Reactions Lyo-ready RPA Kit
A72123 1600 μg Lyo-ready T4 Gene 32 Protein
A72124 120 μg Lyo-ready T4 UvsX Protein
A72125 120 μg Lyo-ready T4 UvsY Protein
A72126 2000 μL Reaction Buffer
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Catalog No. A72127 Supplier Invitrogen™ Supplier No. A72127
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Description

Invitrogen Lyo-ready RPA reagents are designed for fast, sensitive, and specific DNA amplification through isothermal recombinase polymerase amplification (RPA) reactions. Lyo-ready RPA reagents are available in stand-alone and kit formats.

Invitrogen Lyo-ready recombinase polymerase amplification (RPA) reagents are designed for fast, sensitive, and specific DNA amplification through isothermal RPA reactions. Lyo-ready RPA reagents are available in stand-alone and kit formats. Reverse transcriptase can be added to the RPA reaction for amplification of RNA targets. All reagents are glycerol-free, which enables assay lyophilization.

Features of Lyo-ready RPA products

  • Enhanced sensitivity down to 1 copy using optimized (RT-)RPA reaction buffer
  • Robust and specific RNA/DNA amplification
  • Amplification from inhibitor-containing samples
  • Fast target amplification in as little as 20 minutes
  • Ability to easily optimize RPA or RT-RPA reactions

Kit components

The Lyo-ready RPA Kit includes all required components for an RPA reaction:

  • Lyo-ready T4 UvsX Protein—produced from E. coli cells bearing cloned uvsX gene derived from bacteriophage T4
  • Lyo-ready T4 UvsY Protein—a recombination mediator protein that stimulates homologous recombination performed by the UvsX recombinase
  • Lyo-ready Bst DNA Polymerase (Cat. No. A56655)—a proprietary mutant of Bst DNA Polymerase, Large Fragment, with fast reaction speed, increased sensitivity, and tolerance to inhibitors
  • Lyo-ready T4 Gene 32 Protein—a single-stranded DNA binding protein produced from E. coli cells carrying cloned bacteriophage T4 gene 32
  • Optimized reaction buffer, magnesium chloride, and dNTPs

Specifications

Product Type Lyo-ready RPA Kit
Form Liquid
For Use With (Application) RPA, RT-RPA
Polymerase Lyo-ready Bst DNA Polymerase
Quantity 100 Reactions
Research Category Nucleic Acid Amplification
Content And Storage

• Lyo-ready T4 UvsX Protein (1.2 mg/mL), 50 μL
• Lyo-ready T4 UvsY Protein (1.2 mg/mL), 50 μL
• Lyo-ready Bst DNA Polymerase (6 U/μL), 50 μL
• Lyo-ready T4 Gene 32 Protein (16 mg/mL), 50 μL
• 2X RPA Reaction Buffer, 1000 μL
• 280 mM MgCl2, 100 μL
• 10 mM dNTP Mix, 50 μL

Store at -25°C to -15°C.
Optimal Reaction Temperature 42°C
Reverse Transcriptase SuperScript IV Reverse Transcriptase
Shipping Condition Dry Ice
Detection Method End point, real-time, lateral flow
Reaction Time Up to 20 min.
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Frequently Asked Questions (FAQs)

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.
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.

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.

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.
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.

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.

For Research Use Only. Not for use in diagnostic procedures.