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BioReagent,PCR试剂,生物染色剂,用于显微镜,适用于荧光分析 级 提供 ≥95%(HPLC) 纯度,适用于对基线干扰要求严格的色谱和分析工作流程。
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在色谱分析、有机合成和交叉偶联反应领域已被 16 篇同行评审文献引用。
本产品是SYBR Green I 溶于DMSO 的SYBR Green I 1000×溶液。
SYBR Green I 是一种使用便捷、灵敏度高、荧光亮度强的核酸染料。它可以与双链DNA(dsDNA)的双螺旋的小沟区域结合,然后发出绿色荧光。SYBR Green I 在游离状态下,荧光强度比较微弱,一旦与双链DNA结合, 其荧光强度会显著增强。在PCR反应体系中,SYBR Green I 的荧光强度与双链DNA的浓度成正比,PCR产物的增加与荧光强度的升高同步进行。这样我们就可以通过检测荧光强度来对PCR扩增的产物进行定量。SYBR Green I 的最大吸收波长约为497nm,最大发射波长约为520nm,它可以用于qPCR、等温扩增或基因芯片等的荧光定量。
产品优势
(1)荧光信号强:扩增的荧光信号强度高,便于观察实验结果和数据分析;
(2)动态扩增曲线优异:扩增不同浓度的样本均可得到可靠的扩增曲线;
(3)分辨性能好:可精准分辨浓度差异2倍的样本;
(4)出色的稳定性:光稳定性和热稳定性均非常好;
(5)通用性好:绝大多数类型的引物模板均适用;
(6)操作简便:只需在预混液中加入染料即可。
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;然后用ddH₂O 将本产品稀释至100×待 用(例如取20μL SYBR GreenⅠ1000×,则需加180μL的DMSO进行稀释,得到200μL的100× SYBR GreenⅠ)。
2.按下表建立实验体系(仅供参考)
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【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/25μL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 1000× stock solution of SYBR Green I dissolved in DMSO.SYBR Green I is a user-friendly nucleic acid dye featuring high sensitivity and strong fluorescence intensity. It binds to the minor groove of double-stranded DNA (dsDNA) helix and emits green fluorescence. Free unbound SYBR Green I exhibits weak intrinsic fluorescence, while its fluorescence intensity increases dramatically upon binding to dsDNA. In a PCR reaction system, the fluorescence intensity of SYBR Green I is proportional to the concentration of dsDNA; the accumulation of PCR products synchronously elevates the fluorescence signal. Therefore, quantitative analysis of PCR amplicons can be achieved by measuring fluorescence intensity. SYBR Green I has a maximum excitation wavelength of approximately 497 nm and a maximum emission wavelength of around 520 nm. It is applicable to quantitative detection platforms including qPCR, isothermal amplification and DNA microarrays.
Product Advantages
1. Strong fluorescence signal: High signal intensity after amplification, facilitating result observation and data analysis.
2. Excellent amplification curve performance: Reliable amplification curves can be generated for samples with varying template concentrations.
3. Superior discrimination capability: Precisely distinguish samples with a 2-fold concentration difference.
4. Outstanding stability: Excellent photostability and thermal stability.
5. Wide compatibility: Compatible with most primer-template combinations.
6. Simple operation: Only requires adding the dye to the master mix.
Operating Protocol
1. Remove the product and equilibrate to room temperature before use. Mix thoroughly and centrifuge after complete dissolution. Dilute the 1000× stock to a working concentration of 100× with ddH₂O (e.g., add 180 μL DMSO to 20 μL SYBR Green I 1000× to prepare 200 μL 100× SYBR Green I).
2. Prepare the reaction system as shown in the table below (for reference only):
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Notes for Reaction Setup
1. Add an appropriate concentration of reference dye ROX or omit ROX according to your qPCR instrument model.
2. Optimize the system composition appropriately based on the specific Taq DNA polymerase used.
① Prepare sufficient master mix according to the number of test samples, then add template DNA (10 ng per 25 μL reaction system).
② Mix the complete reaction mixture 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 control groups for all qPCR experiments.
2. Wear lab coat and disposable nitrile gloves during handling for personal safety and health protection.
3. This product is for research use only, not for clinical or diagnostic applications.
H341: 怀疑引起遗传缺陷
P405: 密闭存放
P501: 将内容物/容器处理到。。。
P281: 根据需要使用个人防护设备。
P203: 使用前,获取、阅读并遵守所有安全说明。
P318: 如果暴露或担心,请就医。
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| 批号(Lot Number) | 证书类型 | 货号 |
|---|---|---|
| 分析证书 | S1524742 | |
| 分析证书 | S1524742 |
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