SuperScript IV RT-LAMP Master Mix for viral pathogen detection

Accelerate viral pathogen research and surveillance with SuperScript IV RT-LAMP Master Mix

The Invitrogen SuperScript IV RT-LAMP Master Mix is a reverse transcription loop-mediated isothermal amplification (RT-LAMP) based solution for faster and simpler detection of viral pathogens, including SARS-CoV-2, influenza, measles and other pathogens. Our master mix reagents provide maximum flexibility to optimize and accelerate your viral pathogen research and surveillance. 

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Product highlights

  • Faster–viral pathogen detection in as little as 5 minutes with evolved Bst DNA Polymerase 
  • Efficient–one step reaction for reverse transcription of RNA to cDNA with SuperScript IV Reverse Transcriptase
  • Sensitive–greater sensitivity and specificity utilizing RNaseOUT recombinant ribonuclease inhibitor and an optimized buffer 
  • Simpler–streamlined workflow: single tube format, only requires a 65℃ heating block 
  • Flexible–several options for evaluating results, including real-time and endpoint detection methods

Applications

  • Isothermal amplification
  • RNA and DNA target detection
  • Assay development for pathogen detection  
  • Point-of-Care device and field assay development

Benefits of  SuperScript IV RT-LAMP Master Mix

Faster RNA amplification speed for SuperScript IV RT-LAMP Master Mix.
Viral pathogen detection in as little as 5 minutes with evolved Bst DNA Polymerase

Detecting synthetic SARS-CoV-2 using real-time read out

Faster RNA amplification speed for SuperScript IV RT-LAMP Master Mix

SARS-CoV-2 RNA amplification speed. SuperScript IV RT-LAMP Master Mix provides fastest RNA target amplification at constant speed even at low target copy number. Amplification speed was determined using synthetic SARS-CoV-2 RNA, same primer set (from Colorimetric ReadiLAMP™ kit, SARS-CoV-2 Cat. No. A52539) and SYTO 9 green fluorescent nucleic acid stain for real-time detection with Applied Biosystems™ QuantStudio™ 5, 6 Flex and 7 Flex Real-Time PCR Systems. NEB WarmStart® LAMP Kit (DNA & RNA), Optigene GspSSD2.0 Isothermal Mastermix and Lucigen LavaLAMP™ RNA Master Mix were used following the manufacturers’ recommended protocols.The values shown are the average of three experiments. 

SuperScript IV RT-LAMP Master Mix demonstrated robust detection at lower RNA copy numbers, detecting as low as 30 copies in less than 10 minutes.

Detecting synthetic SARS-CoV-2 RNA using real-time read out.

SuperScript IV RT-LAMP Master Mix detects as low as 30 copies in less than 10 mins

Detecting low SARS-CoV-2 RNA copy numbers. SuperScript IV RT-LAMP Master Mix amplifies as low as 30 copies of RNA target in under 10 minutes. Amplification speed was determined using synthetic SARS-CoV-2 RNA, same primer set (from Colorimetric ReadiLAMP™ kit, SARS-CoV-2 Cat. No. A52539) and SYTO 9 green fluorescent nucleic acid stain for real-time detection with Applied Biosystems™ QuantStudio 5, 6 Flex and 7 Flex Real-Time PCR Systems. NEB WarmStart® LAMP Kit (DNA & RNA), Optigene GspSSD2.0 Isothermal Mastermix and Lucigen LavaLAMP™ RNA Master Mix were used following the manufacturers’ recommended protocols.The values shown are the average of three experiments.

SuperScript IV RT-LAMP Master Mix provides several options for evaluating results, including real-time and endpoint methods.

Real-Time Fluorescent RT-LAMP Detection

Real-time fluorescent RT-LAMP detection

Figure 1. Detection of the viral RNA target by the real-time fluorescent RT-LAMP assay. Real-time RT-LAMP was performed using real-time PCR instrument and SYTO™ 9 stain, the reaction time was 60 minutes. Two blue curves demonstrate early amplification signal in positive reactions (< 15min). The dark blue curve represents amplification of the purified clinical RNA, and the light blue curve corresponds to the synthetic viral RNA amplification. The grey curve indicates late amplification signal in the no template control.

Endpoint detection method

Color Change by SYBR Green I Staining and Agarose Gel Electrophoresis

SYBR Green staining 1 on Agarose Gel

Figure 2. Visual detection of the endpoint RT-LAMP reaction results
Upper row:
Color changes produced by SYBR Green I staining. Bright green color (right) corresponds to successful amplification of the target RNA. Orange color (left) indicates no amplification.
Lower row:
Agarose gel verification of RT-LAMP endpoint detection results using a 2% E-Gel agarose gel with SYBR Safe stain. A distinct ladder-like pattern in the gel indicates a positive reaction. Absence of the ladder pattern indicates a negative result. The M lane contains the E-Gel 1 Kb Plus Express DNA Ladder.

SuperScript IV RT-LAMP Master Mix also detects DNA targets, enabling a wider range of applications for your research and surveillance of pathogens.

SuperScript IV RT-LAMP Master Mix also detects DNA targets

DNA target amplification. SuperScript IV RT-LAMP Master Mix can be used to amplify DNA targets and demonstrated superior amplification speeds. Amplification speed was determined using DNA plasmid with SARS-CoV-2 fragment, same primer set (from Colorimetric ReadiLAMP ™ kit, SARS-CoV-2 Cat. No. A52539) and SYTO 9 green fluorescent nucleic acid stain for real-time detection with Applied Biosystems™ QuantStudio 5, 6 Flex and 7 Flex Real-Time PCR Systems. NEB WarmStart® LAMP Kit (DNA & RNA), Optigene GspSSD2.0 Isothermal Mastermix and Lucigen LavaLAMP™ RNA Master Mix were used following the manufacturers’ recommended protocols.The values shown are the average of three experiments.

Faster RNA amplification speed for SuperScript IV RT-LAMP Master Mix.
Viral pathogen detection in as little as 5 minutes with evolved Bst DNA Polymerase

Detecting synthetic SARS-CoV-2 using real-time read out

Faster RNA amplification speed for SuperScript IV RT-LAMP Master Mix

SARS-CoV-2 RNA amplification speed. SuperScript IV RT-LAMP Master Mix provides fastest RNA target amplification at constant speed even at low target copy number. Amplification speed was determined using synthetic SARS-CoV-2 RNA, same primer set (from Colorimetric ReadiLAMP™ kit, SARS-CoV-2 Cat. No. A52539) and SYTO 9 green fluorescent nucleic acid stain for real-time detection with Applied Biosystems™ QuantStudio™ 5, 6 Flex and 7 Flex Real-Time PCR Systems. NEB WarmStart® LAMP Kit (DNA & RNA), Optigene GspSSD2.0 Isothermal Mastermix and Lucigen LavaLAMP™ RNA Master Mix were used following the manufacturers’ recommended protocols.The values shown are the average of three experiments. 

SuperScript IV RT-LAMP Master Mix demonstrated robust detection at lower RNA copy numbers, detecting as low as 30 copies in less than 10 minutes.

Detecting synthetic SARS-CoV-2 RNA using real-time read out.

SuperScript IV RT-LAMP Master Mix detects as low as 30 copies in less than 10 mins

Detecting low SARS-CoV-2 RNA copy numbers. SuperScript IV RT-LAMP Master Mix amplifies as low as 30 copies of RNA target in under 10 minutes. Amplification speed was determined using synthetic SARS-CoV-2 RNA, same primer set (from Colorimetric ReadiLAMP™ kit, SARS-CoV-2 Cat. No. A52539) and SYTO 9 green fluorescent nucleic acid stain for real-time detection with Applied Biosystems™ QuantStudio 5, 6 Flex and 7 Flex Real-Time PCR Systems. NEB WarmStart® LAMP Kit (DNA & RNA), Optigene GspSSD2.0 Isothermal Mastermix and Lucigen LavaLAMP™ RNA Master Mix were used following the manufacturers’ recommended protocols.The values shown are the average of three experiments.

SuperScript IV RT-LAMP Master Mix provides several options for evaluating results, including real-time and endpoint methods.

Real-Time Fluorescent RT-LAMP Detection

Real-time fluorescent RT-LAMP detection

Figure 1. Detection of the viral RNA target by the real-time fluorescent RT-LAMP assay. Real-time RT-LAMP was performed using real-time PCR instrument and SYTO™ 9 stain, the reaction time was 60 minutes. Two blue curves demonstrate early amplification signal in positive reactions (< 15min). The dark blue curve represents amplification of the purified clinical RNA, and the light blue curve corresponds to the synthetic viral RNA amplification. The grey curve indicates late amplification signal in the no template control.

Endpoint detection method

Color Change by SYBR Green I Staining and Agarose Gel Electrophoresis

SYBR Green staining 1 on Agarose Gel

Figure 2. Visual detection of the endpoint RT-LAMP reaction results
Upper row:
Color changes produced by SYBR Green I staining. Bright green color (right) corresponds to successful amplification of the target RNA. Orange color (left) indicates no amplification.
Lower row:
Agarose gel verification of RT-LAMP endpoint detection results using a 2% E-Gel agarose gel with SYBR Safe stain. A distinct ladder-like pattern in the gel indicates a positive reaction. Absence of the ladder pattern indicates a negative result. The M lane contains the E-Gel 1 Kb Plus Express DNA Ladder.

SuperScript IV RT-LAMP Master Mix also detects DNA targets, enabling a wider range of applications for your research and surveillance of pathogens.

SuperScript IV RT-LAMP Master Mix also detects DNA targets

DNA target amplification. SuperScript IV RT-LAMP Master Mix can be used to amplify DNA targets and demonstrated superior amplification speeds. Amplification speed was determined using DNA plasmid with SARS-CoV-2 fragment, same primer set (from Colorimetric ReadiLAMP ™ kit, SARS-CoV-2 Cat. No. A52539) and SYTO 9 green fluorescent nucleic acid stain for real-time detection with Applied Biosystems™ QuantStudio 5, 6 Flex and 7 Flex Real-Time PCR Systems. NEB WarmStart® LAMP Kit (DNA & RNA), Optigene GspSSD2.0 Isothermal Mastermix and Lucigen LavaLAMP™ RNA Master Mix were used following the manufacturers’ recommended protocols.The values shown are the average of three experiments.

SuperScript IV RT-LAMP Master Mix

Product Specifications

Number of steps in protocolSingle tube, one-step  
Target detection time5-15 mins
DNA polymeraseBst DNA polymerase mutant
Reverse transcriptaseSuperScript IV Reverse Transcriptase
Other enzymesRNaseOUT Recombinant Ribonuclease Inhibitor
Dye includedSYTO 9 Green Fluorescent Nucleic Acid Stain
Reaction temperature65℃
Sample typePurified RNA (can be used for DNA)
Target detection validatedSARS-CoV-2, Influenza, Measles
DetectionReal-time or end point
Reactions20, 100, 400 or 1000 reactions
Kit contentsSuperScript IV RT-LAMP Master Mix - 1.25ml SYTO 9 Green Fluorescent Nucleic Acid Stain - 30µL
Kit storage temperature-20℃, protected from light
Regulatory classificationRUO
Catalog Number20 rxn (Cat. No. A51800), 100 rxn (Cat. No. A51801), 400 rxn (Cat. No. A51802), 1000 rxn (Cat. No. A51803)

RT-LAMP Workflow for Assay Research and Development of viral pathogens

Example of typical workflow for assay research and development of viral pathogens using purified RNA samples with 

Invitrogen SuperScript IV RT-LAMP Master Mix

RT-LAMP workflow for Assay Research and Development of viral pathogens

RT-LAMP protocol and application note for stand-alone reagents 

Utilize existing catalog reagents, including the Bsm DNA polymerase, with our RT-LAMP protocol and application note to advance your assay development for viral pathogens, including SARS-CoV-2. The RT-LAMP protocol is also useful as a template for testing other RNA targets with relevant primer sets. Learn more at thermofisher.com/lamp

Lyo-ready enzymes are available for large volume orders email MDxenzymes@thermofisher.com or visit www.thermofisher.com/lyo-ready