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RT² qPCR Master Mixes

Benefits of SABiosciences' SYBR® Green qPCR Master Mixes:

Accuracy: Guarantees single gene-specific products with minimal primer dimers.
Reliability: Maximizes amplification efficiencies.
Convenience: Specially formulated all-in-one mixes that match the instrument in your lab.

To learn more about their performance, see our SYBR Green Master Mix Brochure.

Find the qPCR master mix for the instrument available in your lab:
RT² SYBR Green/ ROX: For all ABI Instrumentation
Cat. No. Size
PA-012
PA-012-12
PA-012-24
PA-012-8
PA-112
For 2 RT² Profiler™ PCR Arrays OR 200 25-µl reactions (2.5 ml)
For 12 RT² Profiler™ PCR Arrays
For 24 RT² Profiler™ PCR Arrays
For 4 pack of 384-well PCR Arrays
2000 25-µl reactions (25 ml)
RT² SYBR Green/ Fluorescein: For all BioRad Instrumentation
Cat. No. Size
PA-011
PA-011-12
PA-011-24
PA-011-8
PA-111
For 2 RT² Profiler™ PCR Arrays OR 200 25-µl
For 12 RT² Profiler™ PCR Arrays
For 24 RT² Profiler™ PCR Arrays
For 4 pack of 384-well PCR Arrays
2000 25-µl reactions (25 ml)
RT² SYBR Green:
For instrumentation not requiring reference dye
Cat. No. Size
PA-010
PA-010-12
PA-010-24
PA-010-8
PA-110
For 2 RT² Profiler™ PCR Arrays OR 200 25-µl
For 12 RT² Profiler™ PCR Arrays
For 24 RT² Profiler™ PCR Arrays
For 4 pack of 384-well PCR Arrays
2000 25-µl reactions (25 ml)

Master Mix quality plays a critical role in the performance

The RT² SYBR® Green qPCR Master Mixes contain all of the optimized reagents and buffers needed for SYBR® Green based real-time polymerase chain reactions in all real-time PCR instrumentation

RT² SYBR® Green qPCR Master Mixes, from the experts in high-performance quantitative PCR, provide:

Accuracy: Guaranteed single gene-specific products with minimal primer dimers
Reliability: Maximized amplification efficiencies
Convenience: Matched to the instrument in your lab with specially formulated all-in-one mixes

 
Performance Protocol Components
 

High Amplification Efficiency over a Wide Dynamic Range:

Brighter SYBR Green Signal Means Greater Sensitivity:

Tighter Control of Polymerase Activity Yields Greater Specificity:

SABiosciences Master Mix

Competitor Master Mix

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Performance Protocol Components
 

Ensure that you do not contaminate the 2X real-time PCR cocktail by dividing into aliquots containing the amount of cocktail necessary for the number of reactions you are preparing each day. The rest of the cocktail should be kept in storage away from any sources of template DNA.

When using RT² qPCR Master Mixes with RT² Profiler™ PCR Arrays, RT² qPCR Primer Assays, Methyl-Profiler PCR Arrays, DNA Methylation qPCR Assays, RT² miRNA PCR Arrays, RT² miRNA qPCR Assays, or ChampionChIP PCR Arrays & Real-time ChIP-qPCR Primer Assays, refer to the appropriate User Manuals for reaction mixtures.

  1. For general PCR purposes, mix the following components in a PCR tube:

    12.5 µl RT² SYBR® Green qPCR Master Mixes
    10.5 µl ddH2O
    1.0 µl template cDNA (up to 250 ng)
    1.0 µl gene-specific 10 µM PCR primer pair stock
    25.0 µl final volume

  2. Recommended real-time thermal cycler program. Please find your product for the correct cycling conditions.
    *NOTE: The 10 min step at 95 ºC is required to activate the HotStart Taq DNA polymerase.
    1. RT² Profiler PCR Arrays & RT² Primer Assays

      Select the Cycling Program appropriate for your Real-Time Instrument:

      Two-Step Cycling Program: 95ºC, 10 min; 40 cycles of (95ºC, 15 sec; and 60ºC, 60 sec); or 

      Extended Two-Step Cycling Program: 95ºC, 10 min; 45 cycles of (95ºC, 15 sec; and 60ºC, 60 sec); or

      Three-Step Cycling Program: 95ºC, 10 min; 40 cycles of (95ºC, 15 sec; 55ºC, 30-40 sec, and 72ºC, 30 sec)

      For the following products, please refer to the corresponding user manuals for the correct cycling programs:

    2. Methyl-Profiler PCR Arrays & DNA Methylation qPCR Assays
    3. RT² miRNA PCR Arrays & miRNA qPCR Assays
    4. ChampionChIP PCR Arrays & Real-time ChIP-PCR Primers
  3. Program the real-time thermal cycler to detect and record the SYBR® Green signal from every reaction at the end of the 60ºC annealing / extension step of each cycle.
  4. Run your instrument's default melting curve program immediately after the above PCR program, and generate a first derivative dissociation curve. No more than one peak should appear in each reaction

    If your instrument does not have a default melting curve program, run the following program instead:

    95ºC, 1 min; 65ºC, 2 min (OPTICS OFF); 65ºC to 95ºC at 2ºC/ min (OPTICS ON).

NOTE: Be sure to visually inspect the plate after the run for any signs of evaporation from any of the wells. If evaporation is observed, make a note of which wells so that you may qualify your data analysis appropriately.

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Performance Protocol Components
 

Please check the kit components immediately after you receive this package. We are not responsible for any missing items not reported within two (2) business days upon receipt.

Contents:

PA-010, PA-011, PA-012: Two (2) tubes
PA-010-12, PA-011-12, PA-012-12: Twelve (12) tubes
PA-010-24, PA-011-24, PA-012-24: Twenty-four (24) tubes
PA-010-8, PA-011-8, PA-012-8: Eight (8) tubes

Each tube contains 1.4 mL of 2X solution, sufficient for one PCR Array or for 100 25-µL reactions

PA-110, PA-111, PA-112: One (1) bottle

Bottle contains 25 mL of 2X solution, sufficient for 2000 25-µL reactions

Storage Conditions:
The kit is shipped on dry ice or cold packs. For long-term storage, keep at 4 ºC. If entire volume is not be used all at once, divide into aliquots and store at 4 ºC. Avoid repeated freezing and thawing.

Shelf Life:
All reagents are stable for 6 months after receipt if stored at the recommended temperature.

NOTICE TO PURCHASER: LIMITED LICENSE

Use of this product is covered by one of more of the following US patents and corresponding patent claims outside the US: 5,079,352, 5,789,224, 5,618,711, 6,127,155, 5,677,152 (claims 1 to 23 only), 5,773,258 (claims 1 and 6 only), 5,407,800, 5,322,770, 5,310,652, 5,994,056, 6,171,785, and claims outside the US corresponding to US Patent No. 4,889,818. The purchase of this product includes a limited, non-transferable immunity from suit under the foregoing patent claims for using only this amount of product for the purchaser's own internal research. No right under any other patent claim (such as apparatus or system claims in US Patent No. 6,814,934) and no right to perform commercial services of any kind, including without limitation reporting the results of purchaser's activities for a fee or other commercial consideration, is conveyed expressly, by implication, or by estoppel. This product is for research use only. Diagnostic uses under Roche patents require a separate license from Roche. Further information on purchasing licenses may be obtained by contacting the Director of Licensing, Applied Biosystems, 850 Lincoln Centre Drive, Foster City, California 94404, USA.

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