Contents

The Imagen ® TaqMan multiplex RT-qPCR Premix is supplied as a ready-to-use 2x reaction mix. The formulation contains, Hotstart Taq DNA polymerase, Fast Reverse transcriptases, dNTPs, MgCl₂, reaction enhancers, and stabilizers.

Storage

• -20 °C
• Protected from light
• Avoid repeated freezing and throwing

Reaction Mix Thawing and Handling

Imagen ® TaqMan multiplex RT-qPCR Premix is delivered in a 2x ready-to-use format. To use the mix, thaw the vial on ice to 4 °C.

Please completely mix the vial and briefly centrifuge to ensure all components are at the bottom of the tube. Store on ice protected from light until ready to use. If using automated liquid handling, let sit at ambient temperature for 10 min to further reduce the viscosity.

Application

• Quantitative 1-step real-time RT-PCR
• Gene expression analysis
• Low copy gene detection

Prepare the qPCR Reaction Mix

1. Mix the Imagen ® TaqMan multiplex RT-qPCR Premix thoroughly but gently until it’s completely homogenous.
2. Prepare the qPCR Reaction Mix for the number of reactions required as shown in table below and plus 10% overage.

3. Vortex the tube to mix the contents thoroughly, then centrifuge briefly to collect the contents at the bottom of the tube. (*Use good pipetting practice to ensure assay precision and accuracy of dispensing.)

4. Add DNA (and nuclease-free water, if needed) to the PCR tubes or wells containing the reaction mix, seal　tubes or wells with flat caps or optically transparent film, and gently vortex to ensure thorough mixing of the reaction components.
5. Program the thermal cycling protocol on the real-time PCR instrument.

6. Load the PCR tubes or plates onto the real-time PCR instrument and start the qPCR run program.
7. When thermal cycling is complete, perform data according to the instructions in the instrument-specific software.