IsoFast™ Hot Start Bst Colour Mix

Art No | PB80.51

IsoFast™ Hot Start Bst Colour Mix

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from 360.00 CHF

IsoFast™ Hot Start Bst Colour reagents combine IsoFast™ Hot Start Bst Polymerase with a pH-based dye for rapid positive/negative screening.

Description

IsoFast™ Hot Start Bst Polymerase Colour and IsoFast™ Hot Start Bst Colour Mix are the most recent and improved development of isothermal amplification technology, designed to deliver unparalleled specificity and speed in DNA amplification processes in order to meet the demands of molecular diagnostics and research.

Fast colour readout for positive/negative testing
AptaLock™ hot start for ultra-sensitive detection of DNA targets
Rapid polymerisation for faster time to results (as little as 10 mins)
Detect down to 3 target copies per μL
Ideal for both cold and room temperature setup
Improved speed and sensitivity for early target detection
High activity at a broad range of temperatures from 55-70°C

Whether you are working with loop-mediated isothermal amplification (LAMP) or other isothermal amplification methods, these IsoFast™ Hot Start Bst Colour reagents guarantees exceptional sensitivity and reliability. The mix harnesses the power of a hot start Bst DNA polymerase enzyme, which ensures minimal background amplification and maximises target specificity. With IsoFast™ Hot Start, your isothermal amplification experiments will be faster, more efficient, and easier than ever, setting new standards for molecular biology techniques in both diagnostic applications and research endeavours.

Hot start is important in isothermal amplification techniques because it helps to prevent non-specific amplification and false-positive results. Isothermal amplification methods, such as loop-mediated isothermal amplification (LAMP), rely on specific primers and enzymes to amplify a target sequence at a constant temperature, typically between 60°C and 65°C. These techniques are widely used in various applications, including molecular diagnostics and pathogen detection.

Material Safety Data Sheet (PDF)

Product manual (PDF)

Why hot start is critical in colourimetric isothermal amplification?

Preventing non-specific amplification in isothermal reactions:

In isothermal amplification, it’s essential to ensure that the amplification process only targets the specific sequence of interest. Non-specific amplification can occur when primers bind to unintended regions of the template or when amplification enzymes prematurely initiate amplification. Hot start techniques involve inhibiting the amplification reaction until the reaction reaches the desired temperature. This prevents the formation of non-specific products during the initial stages of the reaction when the temperature is still below the optimal amplification temperature.

Enhancing isothermal amplification specificity:

By preventing premature amplification, hot start methods increase the specificity of the isothermal amplification reaction. This means that the reaction is less likely to amplify non-target sequences, reducing the risk of false-positive results.

Improving isothermal amplification sensitivity:

Hot start techniques can also improve the sensitivity of isothermal amplification assays. By minimizing non-specific amplification, more of the available reagents are reserved for amplifying the target sequence, increasing the efficiency of the reaction and the detection limit.

Fig-1.-IsoFast-HS-colour-mix-vs-NEB-cold-vs-RT-setup

Isothermal amplification of a target sequence in the scaffolding protein gene from the M13 bacteriophage genome using IsoFast™ Hot Start Bst
Polymerase or IsoFast™ Bst Polymerase in 10x IsoFast™ Colour Buffer A, and NEB WarmStart Colorimetric LAMP 2X Master Mix. A primer mix
consisting of 0.2 μM for F3 and B3 primers, 1.6 μM for FIP and BIP primers and 0.8 μM for LoopF and LoopB primers was used. The total reaction
volume was 25 μL. 8 serial dilutions of M13 ssDNA genome were used, starting with a stock of 0.5 ng/μL and using a dilution factor of 10,
corresponding to the number of genome copies indicated next to the plates. Reaction master mixes and plates were prepared either using cold
blocks (cold Setup) or at room temperature (ambient setup), for approximately 20 min. The reaction was run at 65 °C for 30 minutes. Plates were
then photographed to show the colours obtained at the end of the run. IsoFast™ Bst Polymerase and IsoFast™ Hot Start Bst Polymerase showed a
better sensitivity compared to NEB WarmStart Colorimetric LAMP 2X Master Mix in Cold Setup.

IsoFast™ Hot Start Bst Polymerase allows easy screening for positives even with ambient temperature setup.