Volcano2G DNA polymerase

Volcano 2G DNA polymerase

  • Volcano 2G is an advanced version of the original Volcano DNA polymerase with further improved reverse transcription activity. This enzyme is an engineered, extremely thermostable reverse transcriptase and combined DNA polymerase, obtained through directed, artificial evolution. Volcano2G DNA polymerase facilitates “zero-step” RT-PCRs directly from RNA templates (without an isothermal reverse transcription step), as reverse transcription takes place simultaneously with DNA amplification during the cycled PCR elongation step. This also allows reverse transcription reactions at high temperatures, thus minimizing the problems encountered with strong secondary structures in RNA that melt at elevated temperatures.

  • Volcano2G DNA polymerase contains an hotstart-formulated enzyme blend of an engineered, thermostable DNA polymerase with reverse transcriptase activity and a robust Taq DNA polymerase. The aptamer-based hotstart-formulation is designed to block unspecific DNA polymerase activity at low temperatures (e.g. while pipetting the reaction mixture) but will break away at temperatues above 50-55°C, releasing the the now fully active DNA polymerase.

    Volcano2G DNA polyerase is shipped as a 5 U/µl glycerol stock together with an suitable 5x Volcano reaction buffer optimized for aplicons between 60-300 bp.

    Volcano2G reverse transcriptase can reliably detect RNA targets from ≥ 1 ng of total RNA input. It does not require Mn2+ for optimal activity. However, some assays may be further optimized by the addition of Mn2+ to the reaction (suggested range is 0.5 - 1 mM, Mn2+ not provided).

  • This product is shipped on cool packs. It is recommended to store the product upon arrival at -20°C.

  • RT-PCR activity: Volcano3G RT-PCR Mix is tested for a successful RT-PCR performance. A 151 bp fragment (HPRT1 mRNA) is amplified from human total RNA extract in a real-time PCR cycler and visualized as a single amplified product.

    DNA polymerase activity: Volcano3G DNA polymerase activity is monitored and adjusted to a specific DNA polymerase activity using an artificial DNA template and DNA primer.
    Enzyme concentration is determined by protein-specific staining. Please inquire more information at info@mypols.de for the lot-specific concentration.

Save time with 0-step protocols

Skip the reverse transcription step. Isothermal reverse-transcription steps are not needed. 

Volcano2G is a PCR-active reverse transcriptase and extremely thermostable with a half-life at 95°C of >40 min.

Sensitive RNA detection

Volcano2G DNA polymerase can also be used for real-time PCR, when adding a real-time PCR dye. Volcano2G has a Taq DNA polymerase-like nuclease domain making it also suitable for hydrolysis probe based assays. As demonstrated here, NONO mRNA was detected using a 10-fold dilution series of total RNA by using a probe-based qPCR assay and the zero-step RT-PCR protocol.

Saves time and money

VolcanoCell2G RT-PCR 2x Master mix has a streamlined 'zero-step' workflow that allows you to circumvent laborious purification and extraction steps, saving you both time and money.

Superior sensitivity

VolcanoCell2G RT-PCR 2x achieves detection sensitivities down to single cells. In this example, GAPDH mRNA was detected from a single Lund human mesencephalic (LUHMES) cell following the VolcanoCell2G protocol.

A publication with more examples can be found in the Publications section.

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