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Cat. No.: HY-K0524

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2-Color Tracking SYBR qPCR Master Mix (Universal)

Name: 2-Color Tracking SYBR qPCR Master Mix (Universal)
Brand: MedChemExpress
SKU: HY-K0524
Price: 55.00 USD
Availability: InStock
Rating: 5 (1 reviews)

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MCE 2-Color Tracking SYBR qPCR Master Mix (Universal) is a 2× concentration premixed reagent utilizing a special performance-enhanced Taq DNA polymerase protected via a hot-start activation technique, and optimized qPCR buffer system to perform SYBR Green I based quantitative PCR (qPCR). The 500 rxns is defined as the base specification. All larger sizes correspond to incremental volumes of this base.

Storage :
-20℃, 2 years. Protect from light and avoid repeated freeze-thaw cycles.

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Use of MCE Products

Fluorescent Dye

Strong Fluorescence Signal / Multi-Disciplinary Applications / High Fluorescence Specificity

Description & Advantages

This product contains an appropriate concentration of SYBR Green I, an antibody-modified hot-start Taq DNA polymerase, PCR reaction enhancers, and optimized buffer, which effectively inhibits non-specific PCR amplification, significantly improving PCR amplification efficiency. It is suitable for high-sensitivity Real-Time PCR amplification reactions.

Features of MCE 2-Color Tracking SYBR qPCR Master Mix (Universal)

1. Can be used for rapid Real-Time PCR amplification, yielding good standard curves over a wide quantification range, enabling accurate quantification and detection of target genes with high repeatability and reliability.

2. Using the color-changing effect generated by mixing different dyes, it serves as a pipetting tracer. When the blue SYBR qPCR premix is mixed with the yellow diluent, the solution changes from blue to green, allowing for confirmation of template addition based on the color change. This aids in pipetting large numbers of samples and reduces the probability of errors.

3. Contains a unique reference dye and is compatible with a range of qPCR instruments without the need for additional dyes.

Protocols

1. Preparation of the reaction mixture: prepare the reaction mixture on ice. Three technical replicates are recommended for each sample.

Component	50 μL reaction	20 μL reaction	Final concentration
2-Color Tracking SYBR qPCR Master Mix (Universal)	25 μL	10 μL	1×
Forward Primer (10 μM)	1 μL	0.4 μL	0.2 μM
Reverse Primer (10 μM)	1 μL	0.4 μL	0.2 μM
DNA/cDNA	4 μL	2 μL	/
ddH₂O	19 μL	7.2 μL	/
Total volume	50 μL	20 μL	/

Note: a. A final primer concentration of 200 nM is commonly used and may be optimized within 0.1-1.0 μM according to amplification efficiency and specificity;
b. Recommended template amount: 10-100 ng genomic DNA or 1-10 ng cDNA. Adjust the template amount according to target abundance to keep Ct values preferably within 15-35;
c. It is recommended to prepare a Master Mix without template first, dispense it, and then add template to reduce pipetting error.
d. The formulation contains a two-color tracking system for visual confirmation during reaction setup; no separate ROX Reference Dye is required.

2. qPCR cycling program:

Step	Temperature	Time	Cycles
Initial denaturation	95°C	30 sec	1
Denaturation	95°C	10 sec	40
Annealing/Extension	60°C	30 sec	40
Melting curve	Use the instrument default program	-	-

3. Perform melting-curve analysis after amplification to confirm product specificity. Ramp rates and fluorescence acquisition settings may be adjusted according to the real-time PCR instrument used.