本产品是 MiGreen 分子溶于水的 20× 溶液。MiGreen是一种新型的 AIE 绿色荧光核酸饱和染料，和 SYBR Green I 染料类似，可用于实时荧光定量 PCR（qPCR）等分子生物学实验。MiGreen饱和染料分子在游离状态下，其荧光强度非常微弱，但与双链 DNA（dsDNA）结合后可以发出强烈的绿色荧光。它的光谱信息与 SYBR Green I 类似，结合 dsDNA 的最大激发波长约 496nm，最大发射波长为 526nm，所以MiGreen 饱和染料可以很好地兼容主流 qPCR 仪器。此外，和 SYBR Green I 染料相比，MiGreen 饱和染料有其独特的优势：它对 PCR 反应的抑制更小，这意味着可以使用更高浓度的染料来增强信号，而不会影响反应效率。凭借着其出色的性能，MiGreen 饱和染料可用于 qPCR、HRM、数字 PCR（ddPCR）以及等温扩增等多种核酸检测技术中。
【产品优势】
相较于传统的 SYBR Green I 染料，MiGreen 饱和核酸染料具有以下显著优越性：
(1) 极低的 PCR 抑制性：即使使用较高的染料浓度，MiGreen 饱和染料也不会影响 PCR 反应的效率；
(2) 消除染料重排：MiGreen 饱和核酸染料与双链 DNA 结合时，不会发生染料重排，这使得它可以适用于 HRM 分析；
(3) 高灵敏度：MiGreen 饱和核酸染料可以在 PCR 反应中使用饱和浓度，因此可以获得更高的荧光强度，同时可以提升检测的灵敏度；
(4) 更好的安全性：MiGreen 分子不可以透过细胞膜，因此无法与活细胞内的基因组 DNA 相互作用，因此使用时更加安全，废液处理也更加环保。
(5) 超强的稳定性：MiGreen 分子具有极佳的热稳定性、化学稳定性和光稳定性。它可在 PCR 缓冲液中耐受 95-100℃的高温，也能承受反复冻融，性能不受影响。
【使用方法】
1. 实验前先将本产品取出，置于常温，待其溶解后，混匀离心。
2. 按下表建立实验体系（仅供参考）
| Reagent | Final concentration in the mixture |
|---|
| ddH₂O | Adjust to final volume | | Taq polymerase buffer without magnesium | 1× | | dNTP | 0.2mM each | | MgCl₂ | 2.5mM | | MiGreen | 1× | | Taq DNA polymerase | 1-5 units per reaction | | Each of primers | 0.1-1uM |
|
【注】：1) 请根据不同的仪器类型，不加入或加入合适浓度的参比染料 ROX。
2) 该体系需根据所使用的 Taq 酶的不同进行适当优化。
① 根据检测样本数配置足量的上述反应体系，并加入模板 DNA（10ng/25uL 体系）。
② 将以上体系混匀后分装至 qPCR 管内。
③ 在合适的仪器内启动 qPCR 反应并记录退火或延伸步骤的荧光信号。
【注意事项】
1、做 qPCR 时注意设置阳性和阴性对照；
2、为了您的安全和健康，请穿实验服并戴一次性手套操作；
3、本品仅适用于科研用途。
This product is a 20× aqueous solution of MiGreen molecules.
MiGreen is a novel AIE green fluorescent saturated nucleic acid stain, similar to SYBR Green I, and applicable to molecular biology experiments such as real-time quantitative PCR (qPCR). Free MiGreen saturated stain molecules exhibit extremely weak fluorescence, yet they emit intense green fluorescence upon binding to double-stranded DNA (dsDNA). Its spectral properties are comparable to SYBR Green I, with a maximum excitation wavelength of approximately 496 nm and a maximum emission wavelength of 526 nm when bound to dsDNA, enabling excellent compatibility with mainstream qPCR instruments. Furthermore, MiGreen saturated stain boasts unique advantages over SYBR Green I: it imposes minimal inhibition on PCR reactions, allowing higher stain concentrations to boost signal intensity without compromising reaction efficiency. With outstanding performance, MiGreen saturated stain is compatible with multiple nucleic acid detection technologies, including qPCR, HRM, digital PCR (ddPCR) and isothermal amplification.
Product Advantages
Compared with conventional SYBR Green I stain, MiGreen saturated nucleic acid stain delivers the following prominent benefits:
1. Extremely low PCR inhibition: MiGreen saturated stain does not impair PCR efficiency even at elevated stain concentrations;
2. No dye rearrangement: No dye rearrangement occurs when MiGreen saturated nucleic acid stain binds to dsDNA, making it suitable for HRM analysis;
3. High sensitivity: MiGreen saturated nucleic acid stain can be used at saturating concentrations in PCR systems to generate stronger fluorescence signals and improve detection sensitivity;
4. Enhanced safety: MiGreen molecules cannot penetrate cell membranes and therefore fail to interact with genomic DNA inside live cells, ensuring safer operation and more eco-friendly waste disposal;
5. Excellent stability: MiGreen molecules possess outstanding thermal, chemical and photostability. It withstands high temperatures of 95–100 °C in PCR buffer and repeated freeze-thaw cycles without performance degradation.
Operating Procedures
- Take the product out and equilibrate to room temperature before experiments; mix thoroughly and centrifuge briefly after complete dissolution.
- Prepare the reaction system as shown below (for reference only)
| Reagent | Final concentration in the mixture |
|---|
| ddH₂O | Adjust to final volume | | Taq polymerase buffer without magnesium | 1× | | dNTP | 0.2 mM each | | MgCl₂ | 2.5 mM | | MiGreen | 1× | | Taq DNA polymerase | 1–5 units per reaction | | Each of primers | 0.1–1 μM |
|
Notes: 1. Add reference dye ROX at an appropriate concentration or omit it according to different instrument models.
2. This system requires proper optimization based on the specific Taq enzyme used.
① Prepare sufficient reaction mixture according to the number of test samples, then add template DNA (10 ng per 25 μL reaction system).
② Mix the prepared system thoroughly and aliquot into qPCR tubes.
③ Run the qPCR program on a compatible instrument and record fluorescence signals at the annealing or extension step.
Precautions
1. Set up positive and negative controls during qPCR assays;
2. Wear a lab coat and disposable gloves during operation for personal safety and health protection;
3. This product is for research use only.
ZJ26F0838371
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