IsoFast™ Bst 1-Step Mix

IsoFast™ Bst 1-Step Mix is a dual enzyme system for rapid and sensitive isothermal amplification of RNA targets in one step.

The kit contains IsoFast™ Bst Polymerase, providing strong strand displacement capabilities, together with the highly active modified MMLV RTase Go.

Features

  • Validated for qualitative detection of SARS-CoV-2 nucleic acid
  • Includes strand-displacing IsoFast™ Bst Polymerase in a 2x mix format
  • Supplied with RTase Go plus RNase inhibitor
  • Reaction is carried out at 65˚C
  • Gives rapid and consistent amplification across a wide range of templates 
  • Includes an advanced buffer system for higher yield under difficult conditions
  • Supplied with a fluorescent dye for real-time detection 
  • 30 minute protocol
CAT No.
Product
Size
Price
QTY
PB80.21-01
IsoFast™ Bst 1-Step Mix
100 x 25 μL Reactions
$270.00
PB80.21-05
IsoFast™ Bst 1-Step Mix
500 x 25 μL Reactions
$1,082.00

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More Information

IsoFast™ Bst 1-Step Mix is designed for reverse transcription of target RNA and subsequent isothermal amplification in a single tube. 

Dual enzyme system

The kit utilises IsoFast™ Bst Polymerase for its strong strand displacement activity, enabling DNA synthesis at a constant temperature (65˚C) without the need for thermal cycling. Representing the large fragment of Geobacillus stearothermophilus (formerly Bacillus stearothermophilus) DNA Polymerase, this portion of the protein catalyses the 5’-3’ synthesis of DNA but does not contain the 5’-3’ exonuclease domain.The enzyme is provided in a convenient 2x mix format.

Reverse transcription of target RNA is carried out by the thermostable and extremely active RTase Go, which is blended with RNase inhibitor to prevent degradation of RNA by contaminating RNase.

Rapid and consistent results

Designed for fast amplification, IsoFast™ Bst 1-Step Mix gives rapid and consistent results across a wide range of sample types and is validated for qualitative detection of SARS-CoV-2 nucleic acid (Fig 1 and 2). The buffer chemistry ensures high yield and performance even under difficult conditions, for example when inhibitors are present.

IsoFast™ Bst 1-Step Mix contains all the components for fast and sensitive isothermal amplification of RNA targets, requiring only the addition of primers, template and water. The kit is provided with a separate tube of fluorescent dye to allow real-time detection with any qPCR instrument. 

This product is not suitable for PCR.

Mead DA, McClary JA, Luckey JA, Kostichka AJ, Witney FR, Smith LM. Bst DNA polymerase permits rapid sequence analysis from nanogram amounts of template. Biotechniques. 1991 Jul;11(1):76-8, 80, 82-87.

Applications

  • Multiple displacement amplification
  • Isothermal amplification
  • Reverse Transcription Loop mediated isothermal amplification (RT-LAMP)
  • Molecular diagnostics
  • Field diagnostics

Specifications

IsoFast™ Bst 1-Step Mix

Component

100 Reactions

500 Reactions

IsoFast™ Bst Mix

1 x 1.25mL

4 x 1.6mL

RTase Go

1 x 200µL

1 x 1mL

20x Fluorescent Dye

1 x 125µL

1 x 625µL

IsoFast™ Bst 1-Step Mix

Component

IsoFast™ Bst Mix

RTase Go

20x Fluorescent Dye



100 Reactions

1 x 1.25mL

1 x 200µL

1 x 125µL

500 Reactions

4 x 1.6mL

1 x 1mL

1 x 625µL

Reaction Volume

Storage

25μL

On arrival, products should be stored between -30 and -15°C. If stored correctly the kit will retain full activity for 12 months.

Reaction Volume

25μL

Storage

On arrival, products should be stored between -30 and -15°C. If stored correctly the kit will retain full activity for 12 months.

Documents

FAQs

Can IsoFast™ Bst 1-Step Mix be heat inactivated?

Yes, both IsoFast™ Bst Polymerase and RTase Go can be inactivated by heating the tube at >80°C for 10 minutes.

Can IsoFast™ Bst 1-Step Mix  be used at temperatures other than 65°C?

Yes, the enzyme is active between 57°C and 65°C. We do not recommend using it at temperatures outside this range as the activity will drop.

Can IsoFast™ Bst 1-Step Mix be used for thermal cycle sequencing?

Unfortunately no, as the reaction temperatures are too high for enzyme stability.

Does IsoFast™ Bst Polymerase in IsoFast™ 1-Step Mix have reverse transcriptase activity?

Yes, but it is too low for use in reverse transcription applications. For some primer sets and targets IsoFast™ Bst Polymerase may be used as a single-enzyme (RT/DNA polymerase), but we recommend using the dedicated reverse transcriptase RTase Go as indicated.

How do I use the fluorescent dye?

The intercalating fluorescent dye is provided at a 20x concentration. 1.25μL per 25μL reaction is recommended when monitoring reactions on most real time qPCR instruments. The FAM channel of common real-time fluorimeters can be used to read the dye.

How fast should I expect a result?

Amplification of any target varies depending on several factors including primer design, template quality and quantity. Generally, kits in the IsoFast™ Bst range will achieve positive results between 5 and 30 minutes.

Is IsoFast™ Bst 1-Step Mix supplied with dNTPs?

Yes. Both the buffer and the mix already contain dNTPs, so there is no need to order dNTPs separately.

What components are included in the IsoFast™ Bst 1-Step Mix kit?

The kit consists of the 2x IsoFast™ Bst Mix (including the amplification buffer, dNTPs, MgSO4, IsoFast™ Bst polymerase) and of RTase Go at concentrations optimized for reverse transcription isothermal amplification. The kit also contains one tube of 20x fluorescence dye that binds to dsDNA for real-time detection of products. The user only needs to add primers and target RNA.

When should IsoFast™ Bst Polymerase be the enzyme of choice?

IsoFast™ Bst Polymerase is a DNA polymerase with a very good strand displacement activity. As it has been isolated from the moderate thermophile Bacillus stearothermophilus (recently renamed as Geobacillus stearothermophilus), the temperature optimum of 60-65°C is higher than the one of DNA Polymerase I, Klenow Fragment or the one of phi29 polymerases (37°C), other frequently used strand displacing polymerases.

This is useful in the design of sequencing strategies as well as isothermal amplification technologies. For example, the higher reaction temperature facilitates sequencing through GC rich regions.

More Information

IsoFast™ Bst 1-Step Mix is designed for reverse transcription of target RNA and subsequent isothermal amplification in a single tube. 

Dual enzyme system

The kit utilises IsoFast™ Bst Polymerase for its strong strand displacement activity, enabling DNA synthesis at a constant temperature (65˚C) without the need for thermal cycling. Representing the large fragment of Geobacillus stearothermophilus (formerly Bacillus stearothermophilus) DNA Polymerase, this portion of the protein catalyses the 5’-3’ synthesis of DNA but does not contain the 5’-3’ exonuclease domain.The enzyme is provided in a convenient 2x mix format.

Reverse transcription of target RNA is carried out by the thermostable and extremely active RTase Go, which is blended with RNase inhibitor to prevent degradation of RNA by contaminating RNase.

Rapid and consistent results

Designed for fast amplification, IsoFast™ Bst 1-Step Mix gives rapid and consistent results across a wide range of sample types and is validated for qualitative detection of SARS-CoV-2 nucleic acid (Fig 1 and 2). The buffer chemistry ensures high yield and performance even under difficult conditions, for example when inhibitors are present.

IsoFast™ Bst 1-Step Mix contains all the components for fast and sensitive isothermal amplification of RNA targets, requiring only the addition of primers, template and water. The kit is provided with a separate tube of fluorescent dye to allow real-time detection with any qPCR instrument. 

This product is not suitable for PCR.

Mead DA, McClary JA, Luckey JA, Kostichka AJ, Witney FR, Smith LM. Bst DNA polymerase permits rapid sequence analysis from nanogram amounts of template. Biotechniques. 1991 Jul;11(1):76-8, 80, 82-87.

Applications

  • Multiple displacement amplification
  • Isothermal amplification
  • Reverse Transcription Loop mediated isothermal amplification (RT-LAMP)
  • Molecular diagnostics
  • Field diagnostics

Specifications

IsoFast™ Bst 1-Step Mix

Component

100 Reactions

500 Reactions

IsoFast™ Bst Mix

1 x 1.25mL

4 x 1.6mL

RTase Go

1 x 200µL

1 x 1mL

20x Fluorescent Dye

1 x 125µL

1 x 625µL

IsoFast™ Bst 1-Step Mix

Component

IsoFast™ Bst Mix

RTase Go

20x Fluorescent Dye



100 Reactions

1 x 1.25mL

1 x 200µL

1 x 125µL

500 Reactions

4 x 1.6mL

1 x 1mL

1 x 625µL

Reaction Volume

Storage

25μL

On arrival, products should be stored between -30 and -15°C. If stored correctly the kit will retain full activity for 12 months.

Reaction Volume

25μL

Storage

On arrival, products should be stored between -30 and -15°C. If stored correctly the kit will retain full activity for 12 months.

Documents

FAQs

Can IsoFast™ Bst 1-Step Mix be heat inactivated?

Yes, both IsoFast™ Bst Polymerase and RTase Go can be inactivated by heating the tube at >80°C for 10 minutes.

Can IsoFast™ Bst 1-Step Mix  be used at temperatures other than 65°C?

Yes, the enzyme is active between 57°C and 65°C. We do not recommend using it at temperatures outside this range as the activity will drop.

Can IsoFast™ Bst 1-Step Mix be used for thermal cycle sequencing?

Unfortunately no, as the reaction temperatures are too high for enzyme stability.

Does IsoFast™ Bst Polymerase in IsoFast™ 1-Step Mix have reverse transcriptase activity?

Yes, but it is too low for use in reverse transcription applications. For some primer sets and targets IsoFast™ Bst Polymerase may be used as a single-enzyme (RT/DNA polymerase), but we recommend using the dedicated reverse transcriptase RTase Go as indicated.

How do I use the fluorescent dye?

The intercalating fluorescent dye is provided at a 20x concentration. 1.25μL per 25μL reaction is recommended when monitoring reactions on most real time qPCR instruments. The FAM channel of common real-time fluorimeters can be used to read the dye.

How fast should I expect a result?

Amplification of any target varies depending on several factors including primer design, template quality and quantity. Generally, kits in the IsoFast™ Bst range will achieve positive results between 5 and 30 minutes.

Is IsoFast™ Bst 1-Step Mix supplied with dNTPs?

Yes. Both the buffer and the mix already contain dNTPs, so there is no need to order dNTPs separately.

What components are included in the IsoFast™ Bst 1-Step Mix kit?

The kit consists of the 2x IsoFast™ Bst Mix (including the amplification buffer, dNTPs, MgSO4, IsoFast™ Bst polymerase) and of RTase Go at concentrations optimized for reverse transcription isothermal amplification. The kit also contains one tube of 20x fluorescence dye that binds to dsDNA for real-time detection of products. The user only needs to add primers and target RNA.

When should IsoFast™ Bst Polymerase be the enzyme of choice?

IsoFast™ Bst Polymerase is a DNA polymerase with a very good strand displacement activity. As it has been isolated from the moderate thermophile Bacillus stearothermophilus (recently renamed as Geobacillus stearothermophilus), the temperature optimum of 60-65°C is higher than the one of DNA Polymerase I, Klenow Fragment or the one of phi29 polymerases (37°C), other frequently used strand displacing polymerases.

This is useful in the design of sequencing strategies as well as isothermal amplification technologies. For example, the higher reaction temperature facilitates sequencing through GC rich regions.