AccuAmp Fast SYBR Green qPCR Master Mix/Low ROX
Cat. No. |
Size |
Price |
Quantity |
11016-100 |
100 x 25-µl reactions (1.25 ml) |
$89.00 |
|
11016-400 |
400 x 25-µl reactions (4 x 1.25 ml) |
$309.00 |
|
11016-01k |
1000 x 25-µl reactions (12.5 ml) |
$729.00 |
|
11016-02k |
2000 x 25-µl reactions (25 ml) |
$1359.00 |
Benefits
- High PCR specificity for use with numerous different targets and templates.
- Outstanding sensitivity with early Ct detection.
- High amplification efficiency over a wide dynamic range.
- Faster results with optimized, ready-to-use master mix.
Description
AccuAmp Fast SYBR® Green qPCR Master Mix/ Low ROX is a 2X concentrated, ready-to-use reaction cocktail that contains all components, except primers and template, for rapid real-time PCR on Applied Biosystems 7500, or 7500 Fast instruments. This master mix provides highly specific and sensitive real-time quantification of genomic DNA and cDNA targets using SYBR® Green I, under fast cycling conditions. Since SYBR® Green dye binds to all double-stranded DNA, it is crucial to reduce/ avoid any nonspecific products and primer–dimers generated during amplification. AccuAmp Fast SYBR® Green qPCR Master Mix/ Low ROX promotes highly specific annealing of primers to the PCR template through our unique combination of proprietary buffer, stabilizers, and specially modified hot-start Taq DNA polymerase, which prevents any polymerase activity before heat activation, thereby eliminating amplification of nonspecific products.
Compatible Instrument
- Applied Biosystems 7500
- Applied Biosystems 7500 Fast
- Applied Biosystems ViiA 7
Storage
AccuAmp Fast SYBR® Green qPCR Master Mix/ Low ROX is stable for 1 year when stored at –20°C in a constant-temperature freezer, protected from light. After thawing, mix thoroughly before using.
Quick-Reference Protocol
1. Thaw AccuAmp Fast SYBR Green qPCR Master Mix/Low ROX (2X), template, and primers. Mix the individual solutions.
2. Prepare a reaction mix.
Component |
Volume/Reaction |
Final Concentration |
AccuAmp Fast SYBR Green qPCR |
12.5 µl |
1X |
Forward Primer |
Variable |
300 nM |
Reverse Primer |
Variable |
300 nM |
Template DNA or cDNA |
Variable |
Up to 100 ng/reaction |
Nuclease-free Water |
Variable |
|
Total Reaction Volume |
25 µl |
|
3. Mix the reaction mix thoroughly. After sealing each reaction, centrifuge reaction tubes briefly to avoid bubbles.
4. Program your thermal cycler.
Step |
Temperature &Time |
Notes |
PCR initial activation step |
95°C, 10 min |
Hot-start Taq DNA polymerase is activated by this heating step. |
2-step cycling (40 cycles)* |
a. 95°C, 10 sec (Denaturation) |
Collect fluorescence data at step b. |
Melting curve analysis |
Refer to instrument instructions |
Strongly recommended |
* The primer Tm may be above or below the temperature of annealing/extension. As a starting point, use primers with Tm 60-65ºC.
5. Place the PCR tubes or plates in the real-time cycler, and start the cycling program.
6. Follow instruction provided by the cycler supplier, and perform a melting curve analysis of the PCR product(s).