TaqPath™ ProAmp™ Master Mix, 1 x 10 mL - FAQs

What Applied Biosystems reagents and instruments can I use with TaqMan SNP Genotyping Assays?

The TaqPath ProAmp Master Mix or TaqPath ProAmp Multiplex Master Mix is recommended for TaqMan SNP Genotyping Assays. Additional PCR master mixes that can be used as well, include the TaqMan Universal PCR Master Mix, No AmpErase, TaqMan Universal PCR Master Mix, TaqMan Universal Master Mix II, TaqMan genotyping Master mix, and GTXpress Master Mix. The TaqMan SNP Genotyping Assays can be used on all the real-time PCR instruments from Applied Biosystems, including 7000, 7300, 7500, 7500 Fast, 7700, StepOne, StepOnePlus, 7900HT, ViiA7, and QuantStudio systems.

What is the passive reference dye in the TaqPath ProAmp Master Mix?

The TaqPath ProAmp Master Mix contains ROX dye as the passive reference dye.

Find additional tips, troubleshooting help, and resources within our Real-Time PCR and Digital PCR Applications Support Center.

The amplification plot shows a high level of noise with TaqPath ProAmp Master Mix (Cat. No. A30865). What can cause this?

This could be caused by selecting an incorrect dye as a passive reference. The TaqPath ProAmp Master Mix contains ROX as a passive reference dye. It needs to be confirmed that ROX has been selected rather than MUSTANG PURPLE, which is present in the TaqPath ProAmp Multiplex Master Mix.

Find additional tips, troubleshooting help, and resources within our Real-Time PCR and Digital PCR Applications Support Center.

Why am I getting unexpected results for genotyping experiments using crude lysates with the TaqPath ProAmp Master Mix (Cat. No. A30865)?

The assay may not be compatible with a crude lysate. We recommend that you use purified genomic DNA of high quality.

Find additional tips, troubleshooting help, and resources within our Real-Time PCR and Digital PCR Applications Support Center.

I see small particles in the reaction mix of the TaqPAth ProAmp Master Mix (Cat. No. A30865). Will this cause my reaction to fail?

The small particles seen in the reaction are components of the TaqPath ProAmp Master Mix; they do not interfere with the reaction and do not affect the physical apparatus, including robotics.

Find additional tips, troubleshooting help, and resources within our Real-Time PCR and Digital PCR Applications Support Center.

Does the TaqPath ProAmp Master Mix (Cat. No. A30865) contain UDG/UNG and/or dUTP?

The TaqPath ProAmp Master Mixes contain both dUTP as well as UDG/UNG enzyme for preventing carry-over contamination.

Find additional tips, troubleshooting help, and resources within our Real-Time PCR and Digital PCR Applications Support Center.

Which enzyme/polymerase is used in the TaqPath ProAmp Master Mix (Cat. No. A30865)?

The enzyme used in the TaqPath ProAmp Master Mix is the Dual-Lock Taq DNA Polymerase.

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Has the TaqPath ProAmp Master Mix (Cat. No. A30865) been tested with Fast mode/cycling using a standard 96-well 0.2 mL block?

The TaqPath ProAmp Master Mix has been tested in the Fast as well as standard mode on a 96-well 0.2 mL block. No difference in results is to be expected.

Find additional tips, troubleshooting help, and resources within our Real-Time PCR and Digital PCR Applications Support Center.

Is there a specific protocol for preparing the lysate to be used with TaqPath ProAmp Mastermix (Cat. No. A30865)?

The standard protocols described in the user guides of the DNA Extract All Reagent Kit and MagMAX DNA Multi-Sample Ultra Kit should be followed.

Find additional tips, troubleshooting help, and resources within our Real-Time PCR and Digital PCR Applications Support Center.

Has the tolerance of the TaqPath ProAmp Master Mix (Cat. No. A30865) to the presence of PCR inhibitors been tested?

Hematin, heparin, IgG, EDTA and citrate have been tested as inhibitors of PCR with the TaqPath ProAmp Master Mix.

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Have you tested the TaqPath ProAmp Master Mix (Cat. No. A30865) on any other type of sample besides blood, dried blood on filter paper, or buccal swabs?

Besides blood, dried blood on filter paper, and buccal swabs, we have tested the TaqPath ProAmp Master Mix on mouse tails, and it worked well in this in-house testing.

Find additional tips, troubleshooting help, and resources within our Real-Time PCR and Digital PCR Applications Support Center.

Can the TaqPath ProAmp Master Mix with ROX (Cat. No. A30865) be used on the 7900HT instrument? Which thermal cycling conditions should be used?

Yes, the TaqPath ProAmp Master Mix can be used on the 7900 HT instrument. Please see the complete protocol here.

Find additional tips, troubleshooting help, and resources within our Real-Time PCR and Digital PCR Applications Support Center.

Can I use TaqPath ProAmp Master Mix (Cat. No. A30865) for CastPCR/TaqMan Mutation Detection Assays?

We have not tested the TaqPath ProAmp Master Mix for use with CastPCR/TaqMan Mutation Detection Assays. The TaqMan Genotyping Master Mix should be used for CastPCR.

Find additional tips, troubleshooting help, and resources within our Real-Time PCR and Digital PCR Applications Support Center.

What is the initial denaturation/enzyme activation time in the Genotyping run method and the copy number variation protocol using the TaqPath ProAmp Master Mix (Cat. No. A30865)?

The activation time for Genotyping protocol with the TaqPath ProAmp Master Mix is 5 minutes. Copy Number Variation (CNV) assays have a 10-minute activation time. It also ensures that the genomic DNA is fully denatured prior to the cycling stage. Please see the complete protocol here.

Find additional tips, troubleshooting help, and resources within our Real-Time PCR and Digital PCR Applications Support Center.

What are the benefits of TaqPath ProAmp Master Mixes (Cat. No. A30865)?

The benefits of the TaqPath ProAmp Master Mix are as follows:

• TaqPath is manufactured in an ISO 13485-certified facility and labeled “For Laboratory Use”.
• TaqPath raw materials, processes, and specifications are tighter than RUO reagents - giving more confidence in lot-to-lot consistency.
• 72 hours pre-PCR benchtop stability enabled by a proprietary Dual-Lock Taq DNA hot-start mechanism.
• TaqPath master mixes are both Fast and Standard cycling compatible.
• TaqPath master mixes are optimized for highly multiplexed applications.

I would like to order a TaqMan SNP Genotyping Assay. Does the kit include everything I need to perform my experiment or do I need to purchase reagents separately?

Each predesigned TaqMan SNP Genotyping Assay is delivered in a single tube consisting of two differentially labeled allele-specific TaqMan MGB (minor groove binding) probes and a pair of PCR primers. For more information, please see the following link: https://www.thermofisher.com/us/en/home/life-science/pcr/real-time-pcr/real-time-pcr-applications/genetic-variation-analysis-using-real-time/snp-genotyping-with-real-time-pcr.html

Besides the TaqMan SNP Genotyping Assay, you would need to purchase a Genotyping Master Mix and nuclease-free water. A list of compatible master mixes can be found in the following link: https://assets.thermofisher.com/TFS-Assets/LSG/manuals/MAN0009593_TaqManSNP_UG.pdf.

Find additional tips, troubleshooting help, and resources within our Gene Expression Analysis & Genotyping Support Center.