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[DESCRIPTION]
GoTaq® qPCR Master Mix(a,b) is a reagent system for quantitative PCR (qPCR). The system contains a new fluorescent DNA-binding dye that often exhibits greater fluorescence enhancement upon binding to double-stranded DNA (dsDNA) than SYBR. Green I.
 
GoTaq® qPCR Master Mix is provided as a simple-to-use, stabilized 2X formulation that includes all components for qPCR except sample DNA, primers and water. This formulation, which includes a proprietary dsDNA-binding dye, a low level of carboxy-X-rhodamine (CXR) reference dye (identical to ROX™ dye), GoTaq. Hot Start Polymerase, MgCl2, dNTPs and a proprietary reaction buffer, produces optimal results in qPCR experiments. A separate tube of CXR Reference Dye is included for use with instruments that require a higher level of reference dye than that in the GoTaq® qPCR Master Mix.
 
Advantages of the GoTaq® qPCR Master Mix
 
Dye: The proprietary dye provides brighter dsDNA-dependent fluorescence than SYBR. Green I, with less PCR inhibition than SYBR® Green. The dye enables efficient amplification, resulting in earlier quantification cycle (Cq) values and an expanded linear range using the same filters and settings as SYBR® Green I. The CXR reference dye can be detected using the same filters and settings as those used for ROX™ dye.
 
Quantification cycle is formerly known as cycle threshold (Ct).
 
Polymerase/Buffer Formulation: GoTaq® Hot Start Polymerase contains full-length Taq DNA polymerase bound to a proprietary antibody that prevents polymerase activity at room temperature. Thermal activation is achieved by incubating the assembled reaction at 95°C for 2 minutes. The proprietary polymerase/buffer formulation accommodates extended cycle numbers (45–50 cycles) and is compatible with thermal cycling programs that require extended activation (95°C for 10 minutes).
 
Performance: You can expect reliable performance with minimal lot-to-lot variation: efficient, sensitive and linear qPCR amplification over a wide dynamic range.
 
GoTaq® qPCR Master Mix Protocol
 
If you are currently performing dye-based qPCR, the GoTaq® qPCR Master Mix can simply be substituted for your current master mix. For consistency within an experimental set, prepare a sufficient volume of reaction mix without template DNA for the DNA standard reactions and experimental sample reactions. The protocol for a 50μl reaction is outlined below. Component volumes may be scaled as appropriate. This protocol assumes that 20% of the reaction volume is DNA template (e.g., 10μl of DNA template added to 40μl of reaction mix). If the volume of DNA template is more or less than 10μl, adjust the volume of Nuclease-Free Water accordingly so that the final reaction volume is 50μl.
 

[GUIDELINES]
<p><strong>Storage Conditions</strong></p>
<p>GoTaq® qPCR Master Mix is shipped at –20°C. Upon arrival, store all components at –20°C, protected from light. For immediate use, components may be stored at 2–8°C, protected from light, for up to 3 months.</p>
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<p><strong>Spectral Properties</strong></p>
<p>The proprietary dye in the GoTaq® qPCR Master Mix has spectral properties similar to those of SYBR® Green I: Excitation at 493nm and emission at 530nm. Instrument optical settings established for SYBR® Green I assays should be used with GoTaq® qPCR Master Mix. The CXR reference dye has the same spectral properties as ROX™: Excitation at 580nm and emission at 602nm. Use the instrument settings for ROX™ dye for reactions containing GoTaq® qPCR Master Mix.</p>
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<p><strong>Magnesium Chloride Concentration</strong></p>
<p>The MgCl<sub>2</sub> concentration of the GoTaq® qPCR Master Mix has been determined to be optimal for performance. If desired, the MgCl<sub>2</sub> concentration may be adjusted using a PCR-grade stock solution (not provided).</p>
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<p><strong>Instrument Compatibility</strong></p>
<p>GoTaq® qPCR Master Mix can be used with any real-time instrument capable of detecting SYBR® Green I or FAM™ dye. GoTaq® qPCR Master Mix contains a low level of CXR reference dye. If you are using any of the following instruments, supplement the GoTaq® qPCR reaction mix with 0.5μl of CXR Reference Dye per 50μl reaction.</p>
<ul>
<li>Applied Biosystems 7000 Sequence Detection System</li>
<li>Applied Biosystems 7300 Real-Time PCR System</li>
<li>Applied Biosystems 7700 Sequence Detection System</li>
<li>Applied Biosystems 7900HT Real-Time PCR System</li>
</ul>

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