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Volcano3G® RT-PCR Probe 2x Master Mix (+low ROX)

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Volcano3G® RT-PCR Probe 2x Master Mix (+low ROX)

$110.00

Volcano3G® RT-PCR Probe 2x Master Mix contains all components required for general RT-qPCR
– optimized reaction mix for sensitive and reliable results
– engineered, truly thermostable Taq DNA polymerases with reverse transcriptase activity
– fast start function due to a hotstart aptamer formulation which prevents unspecific amplification at lower temperatures (<57°C)
– Pipetting aid in form of a blue dye for better visibility during aliquoting into well-plates
– included lysis function*

* = A sample extraction and sample lysis is not necessary as immediate hot PCR cycling conditions can brake cell- and virus membranes. Please see our published references to this product.

Note: Volcano3G® RT-PCR Probe 2x Master Mix is a unique master mix. If you are using it for the first time, we recommend to re-establish the PCR protocol. Primers should be re-designed for higher melting/annealing points (above 65°C) and additionally, RT-PCRs with temperature gradients are performed to establish the best working PCR protocol for your assay with Volcano3G® RT-PCR Probe 2x Master Mix.

This product is for research use only. Please get in touch with us if you need it for diagnostic or commercial purposes.

SKU: myPOLS 6101Hi-S Categories: ,

Description

RT-PCR becomes easier once again

With Volcano3G RT-PCR master mixes, RT-PCR becomes easier once again. Volcano3G RT-PCR master mixes enable amplification of RNA target sequences with quick and easy PCR protocols, even including 0-step amplification.

zero-step RT-PCRs

Be quick! No isothermal reverse transcription step is needed:  A fast start function due to a hotstart aptamer and the  engineered, truly thermostable Taq DNA polymerase with reverse transcriptase activity allows immediate cycling.

Even directRT-PCRs are possible like this: Sample extraction and sample lysis is not necessary as immediate hot PCR cycling conditions can brake cell- and virus membranes. Please see our published references to this product.

Volcano2G Volcano3G comparison

Superior Performance

Volcano3G is the next generation of our Volcano2G with an improved performance. It offers a variety of applications for a precise and rapid qPCR. It is applicable with common thermal cycling protocols and “zero-step” RT-PCR protocols direct from RNA. As demonstrated here, artificial HPRT1 mRNA was detected in a 10-fold dilution series by using a probe-based qPCR assay and a common thermal RT-PCR protocol

Improved Sensitivity

Volcano3G shows an improved detection of mRNA down to 500 total copies, which is clearly distinguishable from the No Template Control (NTC). The brighter fluorescent signal provided by the Volcano 3G Master Mix enables an easy and precise quantification. Volcano3G RT-PCR mix showed to be more sensitive than our Volcano2G RT-PCR mix by a factor of 10. Here, artificial HPRT1 mRNA was detected down to 500 copies with Volcano3G RT-PCR 2x Master mix that contains a sensitive DNA binding dye with negligible polymerase inhibition.

Melt your DNA, not your brain

Volcano3G RT-PCR 2x Master mix with GreenDye is suitable for high resolution melting (HRM) experiments, allowing you to quickly identify unspecific amplicons and thus save time and effort when optimizing your PCR.

Additional information

Applications

– Rapid detection and identification of RNA & DNA targets
– Reverse transcription qPCRs (RT-qPCRs)*
– Direct from cell RT-PCRs*
– qPCRs

*Please see for example our publication regarding
Corona virus (SARS-CoV-2, COVID-19) detections from crude oral swabs: (Detection of SARS-CoV-2 from raw patient samples by coupled high temperature reverse transcription and amplification)
or qPCRs from Cell cultures: (Open access: Reverse-transcription quantitative PCR directly from cells without RNA extraction and without isothermal reverse-transcription: a ‘zero-step’ RT-qPCR protocol)

Variants of Volcano3G® RT-PCR 2x Master Mix are also available with a DNA binding dye (GreenDye), with or without passive reference dye (ROX)(#6301/#6401)
Quality

Volcano3G® RT-PCR Probe 2x Master Mix is tested for a successful, reproducible RT-qPCR performance. Enzyme concentrations are determined by protein-specific staining.

The activities of the enzymes are monitored and adjusted to a specific DNA polymerase activity using a synthetic DNA template and DNA primer.

Additionally, a 151 bp fragment (HPRT1 mRNA) is amplified from an in-vitro transcribed RNA strand by a 0-step real-time RT-qPCR protocol using hydrolysis probes. The linearity of amplification over a specified serial dilution is demonstrated.

No human RNA and DNA as well as no nuclease contaminations is detected.

This product is for research use only but it can also be used for diagnostic or commercial purposes. Please get in touch with us for more information.
Downloads

– Download Volcano3G RT-PCR Probe 2x Master Mix Manual

– Link to publication using Volcano3G for detection of the Corona virus without sample extraction: Detection of SARS-CoV-2 from raw patient samples by coupled high temperature reverse transcription and amplification
https://journals.plos.org/plosone/article?id=10.1371/journal.pone.0241740
FAQs

Please contact us if you have questions for this product.
As Volcano3G is a very unique mastermix, it is not trivial to get started. We have already established PCRs for other customers and offer this service for free to you too.

How do I have to store the mix?C
– Keep at -20°C and minimize the number of freeze-thaw cycles by storing the product in aliquots. For a day-to-day use, we recommend keeping an aliquot at 4°C.

Can I use the mix with crude samples for direct PCRs?
Yes this is possible in general. We have for example tested Volcano with cells and cell-lysates as well as unpurified oral swab samples. More information can be found in the download section of this product.

How stable is the mix?
Volcano3G® RT-PCR Probe 2x Master Mix is shipped on cool-packs around the world. It has been stress-tested at long time periods at elevated temperatures (up to 40°C) without any loss of detectable activity. However we recommend to follow our storage recommendations-

Can I simply replace my currently used RT-PCR mix with your Volcano3G® RT-PCR Probe 2x Master Mix?
No. We recommend to re-establish the PCR protocol. Primers should be re-designed for higher melting/annealing points (above 65°C) and additionally, RT-PCRs with temperature gradients are performed to establish the best working PCR protocol for your assay with Volcano3G.
Catalog Number

#6201HiS