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Home > Products > PCR Home > RT² SYBR® Green qPCR Mastermix
RT² SYBR® Green qPCR Mastermix
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The RT² SYBR® Green qPCR Mastermixes contain all of the optimized reagents
and buffers needed for SYBR® Green based real-time polymerase chain
reactions in all real-time PCR instrumentation including ABI® Bio-Rad®
Stratagene® Roche, and Eppendorf® Each Mastermix includes real-time PCR
buffer, a high-performance HotStart DNA Taq polymerase, nucleotides, and
ROX® or Fluorescein reference dye if and when needed to normalize a
specific instruments' optics. Simply add the Mastermix to PCR tubes along
with your template and primers. The chemically-modified and tightly
controlled HotStart enzyme uniquely provides more accurate SYBR® Green
results by preventing the amplification of primer dimers and other
non-specific products. These RT² SYBR® Green qPCR Mastermixes are best
suited for real-time PCR applications using SYBR® Green based detection
like the RT² Profiler™, RT² miRNA, EpiTect ChIP and EpiTect Methyl qPCR Arrays and qPCR Assays from SABiosciences.
RT² SYBR® Green qPCR Mastermixes, 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
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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 Mastermixes with RT² Profiler™ PCR Arrays, RT²
qPCR Primer Assays, EpiTect Methyl qPCR Arrays, EpiTect Methyl qPCR
Assays, RT² miRNA PCR Arrays, RT² miRNA qPCR Assays, or EpiTec ChIP qPCR
Arrays & EpiTec ChIP qPCR Primer Assays, refer to the appropriate
User Manuals for reaction mixtures.
- For general PCR purposes, mix the following components in a PCR tube:
12.5 µl RT² SYBR® Green qPCR Mastermix
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
- 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.
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RT² Profiler PCR Arrays & RT² Primer Assays
Use Two-Step Cycling Program For:
ABI® 7000, 7300, 7500, 7700, 7900HT
Stratagene Mx3000, Mx3500, Mx4000p
Eppendorf® Mastercycler® ep realplex with ROX® filter set
95ºC, 10 min; 40 cycles of (95ºC, 15 sec; and 60ºC,
60 sec)
Use Three-Step Cycling Program For:
ABI® StepOnePlus™
Cepheid SmartCycler®
All Other
Instruments
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:
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EpiTectMethyl PCR Arrays & DNA Methylation qPCR Assays
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RT² miRNA PCR Arrays & miRNA qPCR Assays
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EpiTectChIP PCR Arrays & Real-time ChIP-PCR Primers
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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.
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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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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:
| Catalog # |
Contents |
| 330520 |
Two (2) tubes each containing 1.25 ml of 2X solution and
enough for 100 standard 25-µl reactions
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| 330522 |
Twelve (12) tubes each containing 1.25 ml of 2X solution
and enough for one PCR Array
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| 330523 |
Twenty-four (24) tubes each containing 1.25 ml of 2X
solution and enough for one PCR Array
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| 330521 |
Eight (8) tubes each containing 1.25 ml of 2X solution and
enough for 4 384-well PCR Arrays
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| 330529 |
One (1) bottle containing 25 ml of 2X solution and enough
for 2000 standard 25-µl reactions |
Storage Conditions:
The kit is shipped on dry ice or cold packs. For long-term storage, keep at -20
ºC. If entire volume is not be used all at once, divide into aliquots and
store at -20 ºC. Avoid repeated freezing and thawing.
Shelf Life:
All reagents are stable for 6 months after receipt if stored at the recommended
temperature.
SYBR® is a registered trademark of Molecular Probes, Inc.; SmartCycler®
is a registered trademark of Cepheid; LightCycler® is a registered trademark
of Roche Diagnostics; ABI® is a registered trademark of Applera Corporation;
FAM™ is a trademark of Applera Corporation; ROX™ is a trademark of Applera
Corporation; Bio-Rad® and iCycler® are registered trademarks of Bio-Rad
Laboratories; Chromo4™ iQ™, MyiQ™ and Opticon™ are trademarks of Bio-Rad
Laboratories; Stratagene® is a registered trademark of Stratagene Corporation;
Eppendorf® and Mastercycler® are registered trademarks of Eppendorf AG; RT² and
RT² Profiler™ are trademarks of SABiosciences Corporation.
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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