All products have special prices for bulk purchase, please contact for more details if required.

Cat. No.: SGQU-N-1 (for 1ml, No ROX)

Cat. No.: SGQU-N-5 (for 5ml, No ROX)

Cat. No.: SGQU-N-25 (for 25ml, No ROX)

Cat. No.: SGQU-L-1 (for 1ml, Low ROX)

Cat. No.: SGQU-L-5 (for 5ml, Low ROX)

Cat. No.: SGQU-L-25 (for 25ml, Low ROX)

Cat. No.: SGQU-H-1 (for 1ml, High ROX)

Cat. No.: SGQU-H-5 (for 5ml, High ROX)

Cat. No.: SGQU-H-25 (for 25ml, High ROX)

Description

SYBR Green qPCR Mix (2X, UDG) is a state-of-the-art, high-quality carryover prevention premix tailored for real-time fluorescence quantitative PCR, also referred to as qPCR or real-time PCR. This mix is designed for ultra-sensitive specific quantification of cDNA and genomic DNA. It contains a precise blend of high-quality UDG enzyme and dUTP, effectively eliminating contamination issues from PCR amplification that can result in false positives or lower CT values. Typically referred to as the dye method due to the use of SYBR Green, this technique can also be named the carryover prevention dye method.

UDG (Uracil-DNA Glycosylase), also known as UNG (Uracil-N-glycosylase), catalyzes the hydrolysis of the N-glycosidic bond between the uracil (dU) base in DNA and deoxyribose, releasing free uracil. It is mainly used to mitigate product contamination issues during PCR amplification. Its anti-contamination mechanism involves incorporating dUTP into DNA during PCR, producing dU-containing amplified products. In subsequent PCR reactions, the UDG enzyme selectively cleaves dU-containing single or double-stranded DNA, minimizing negative effects from previous PCR product contaminations.

The SYBR Green qPCR Mix (2X, UDG) utilizes SYBR Green I dye, which binds to the minor groove of double-stranded DNA (dsDNA). The fluorescence of free SYBR Green I is relatively weak, but intensifies upon binding to dsDNA. This enables quantification of the amount of dsDNA produced during PCR based on fluorescence intensity.

The included Hotstart Taq DNA Polymerase in this product is of premium quality and is heat-activated. It's complexed with an antibody that inhibits the Taq enzyme's DNA polymerase activity, effectively preventing non-specific annealing of primers and template DNA or primer dimerization at lower temperatures. During the PCR's denaturation step, the antibody is deactivated by heat, ensuring the Taq enzyme's activity is only released post-denaturation. This significantly enhances the PCR reaction's specificity, sensitivity, and quantification accuracy.

This product encompasses all generic components, including Hotstart Taq DNA Polymerase, UDG enzyme, PCR Buffer, dNTPs, dUTP, SYBR Green I fluorescent dye, stabilizers, and magnesium ions, making operations simpler and usage more convenient. Users only need to supply primers, sample DNA, and deionized water.

ROX Dye

The role of ROX dye is to correct fluorescence fluctuations unrelated to PCR, thereby minimizing inter-well variability. This variation might be caused by factors such as pipetting errors and sample evaporation. Different quantitative PCR instruments have varying requirements for ROX. Depending on the specific instrument in use, one should choose a SYBR Green qPCR Mix (2X, UDG) containing either a high concentration of ROX (High ROX), a low concentration of ROX (Low ROX), or no ROX. Typically, the SYBR Green qPCR Mix containing a high concentration of ROX can also be used for fluorescence quantitative PCR instruments that don't require ROX or require a low concentration of ROX.

Storage

Precautions

• Before use, ensure the entire tube of reagent is fully thawed. Invert and gently mix before using. During mixing, avoid creating bubbles.
• Pay attention to the annealing temperature of primers. When the annealing temperature is <60ºC, it is recommended to use a three-step PCR amplification.
• This product contains SYBR Green I fluorescent dye. Avoid exposure to strong light when storing this product or setting up PCR reactions to minimize fluorescence quenching issues.
• For amplification fragments exceeding 350bp or with a high GC content, it is suggested to extend the elongation time to 60 seconds or use a three-step method to enhance amplification efficiency.
• Tests show that thawing and refreezing this product up to 10 times does not significantly affect its performance. However, avoid repeated freeze-thaw cycles as they might degrade the product's performance.
• qPCR detection is ultra-sensitive. Although this product has excellent contamination prevention, areas where PCR reactions are set up should still avoid contamination from potential amplification products. PCR products should be sealed and discarded to prevent contaminating the experimental environment with highly concentrated PCR products.
• This product is for scientific research use by professionals only. It is not for clinical diagnosis or treatment, not for use in food or drugs, and should not be stored in ordinary residences.
• For your safety and health, wear a lab coat and use disposable gloves when handling the product.

Related:

• Universal Probe One-Step RT-qPCR Kit
• Probe One-Step qRT-PCR Kit (20X, UDG)
• Multiplex Probe qPCR Mix (2X, UDG)
• Probe qPCR Mix (2X, UDG)
• SYBR Green One-Step qRT-PCR Kit (20X, UDG)
• SYBR Green qPCR Mix (2X, UDG)

Only for research and not intended for treatment of humans or animals

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SBS Genetech is a long-term sponsor of Cold Spring Harbor Laboratory