Ready-to-use Universal qPCR Reference Dye (100×)是一款在实时荧光定量PCR (qPCR)中使用的归一化校正试剂，是通过荧光信号归一化消除实验系统误差。本产品优势在于采用独特长斯托克位移设计，作为5-ROX/6-ROX染料的理想替代品，能够在不同类型的qPCR仪器上实现高效的荧光信号归一化，而且为即用型预混体系，使用时仅需按比例添加至反应体系，即可提供稳定基线无需调整染料浓度，简化操作流程，提高了实验数据的准确性和重复性。本产品应用于qPCR及逆转录-定量PCR (RT-qPCR)的信号归一化与实验优化
市场上现有的5-ROX/6-ROX染料在实时荧光定量PCR中用于荧光信号的归一化，但由于不同仪器平台的光学设计差异，导致在不同仪器上需要使用不同浓度的染料。本产品可解决传统参比染料仪器适配性差、操作繁琐的缺点，无需重复校正热循环仪，为科研人员提供高效、通用、精准的信号归一化、可靠的定量结果解决方案。仅限体外科研用途 不得用于临床诊断或治疗。
注：效能等同Thermo Fisher ROX参比染料
应用范围：实时定量PCR参比染料
产品特点：
1、适配性极强: 无需区分qPCR仪器类型，适用于所有需要或不需要ROX校准的实时荧光定量PCR扩增仪器。
2、 高效稳定基线: 为多重定量PCR和RT-PCR提供稳定的基线。
3、荧光信号归一化: 独特长斯托克位移设计，为qPCR及RT-PCR提供稳定基线，有效消除系统误差。
4、 简化操作: 即用型预混体系，仅需按比例添加至反应体系，无需额外优化浓度，避免反复调试参数。
产品参数：
Ex/Em: 578/604 nm
产品组分：
| 组分 | 2000 T | | A. 100× Ready-to-use Universal qPCR Reference Dye | 1 mL |
|
注：2000 T为试剂盒为试剂盒进行50 μL反应体系的总数。
注意事项：
1、使用前请充分摇匀，避免反复冻融。建议首次使用时将产品分装至小管中，并储存在-20 ℃。
2、荧光染料易受光照影响而淬灭，请务必避光操作，以保持染料稳定性。
3、本产品仅限于科研用途并且不得存放于普通住宅内。
4、为了您的安全和健康，请遵循您所在常规实验室安全规定。
使用说明：
一、实验前准备
1、试剂准备：
将试剂盒中的组分A (100× Ready-to-use Universal qPCR Reference Dye)以及qPCR/RT-PCR反应体系相关试剂，从储存条件 (如-20 ℃或4 ℃)取出，放置于冰上或冰板上。
注：注意避光。
2、仪器准备：qPCR仪
二、 操作步骤
准备定量 PCR/RT-PCR 反应体系：
1、根据PCR试剂系统提供的方案，设置定量 PCR/RT-PCR 反应体系。对于多个反应，首先组装成主混合物，确保混合均匀。
2、添加组分A (100× Ready-to-use Universal qPCR Reference Dye): 在每个50 μL的PCR反应体系中加入0.5 μL的组分A (100× Ready-to-use Universal qPCR Reference Dye)。
3、混匀离心: PCR反应体系震荡混匀后瞬时离心。
注：(1) 根据实验需求参比染料体积可做调整。
(2) 瞬时离心后不能有气泡产生，管壁上不能有液体。
(3) 注意避光。
| Instrument | 50 µL PCR Reaction | Final Concentration | | Bio-Rad CFS 系列 | 0 | 0 | | Roche Light Cyler 系列 | 0 | 0 | | Eppendorf Mastercyler ep realplex 系列 | 0 | 0 | | Qiagen/Corbelt Rotor-Gene 系列 | 0 | 0 | | Takara Thermal Cyler Dice 系列 | 0 | 0 | | ABI PRISM 7000/7300/7700/7900HT and 7900HT Fast system, ABI Step One, ABI Step One Plus 系列 | 0.5 µL | 1× | | ABI 7500/7500 Fast, ViiA7，QuantStudio, Stratagene Mx3000P/3005P/x4000, MJ Research 系列 | 0.5 µL | 1× | | Chromo4, Opticon (II), Corbett Rotor Gene 3000 系列 | 0.5 µL | 1× |
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Product Introduction
The Ready-to-use Universal qPCR Reference Dye (100×) is a normalization calibration reagent for real-time fluorescent quantitative PCR (qPCR), which eliminates experimental systematic errors via fluorescence signal normalization. This product features a unique long Stokes shift design, making it an ideal alternative to 5-ROX/6-ROX dyes. It enables efficient fluorescence signal normalization across various qPCR instrument platforms. As a ready-to-use premixed formulation, it only needs to be added to the reaction system in proportion for use, delivering a stable baseline without the need for dye concentration adjustment. This simplifies the experimental workflow and improves the accuracy and repeatability of experimental data. The product is applicable for signal normalization and experimental optimization in qPCR and reverse transcription-quantitative PCR (RT-qPCR).
Existing 5-ROX/6-ROX dyes on the market are used for fluorescence signal normalization in real-time fluorescent quantitative PCR. However, differences in the optical design of various instrument platforms necessitate the use of different dye concentrations for different instruments. This product addresses the drawbacks of traditional reference dyes, such as poor instrument compatibility and cumbersome operation, and obviates the need for repeated calibration of thermal cyclers. It provides researchers with a solution for efficient, universal and accurate signal normalization, as well as reliable quantitative results. For in vitro research use only. Not for clinical diagnosis or treatment.
Note: Equivalent in performance to Thermo Fisher ROX Reference Dye.
Application Scope
Reference dye for real-time fluorescent quantitative PCR
Product Features
1. Exceptional compatibility: No need to distinguish between qPCR instrument types; suitable for all real-time fluorescent quantitative PCR amplification instruments that require or do not require ROX calibration.
2. Highly stable baseline: Delivers a stable baseline for multiplex quantitative PCR and RT-PCR.
3. Reliable fluorescence signal normalization: The unique long Stokes shift design maintains a stable baseline for qPCR and RT-PCR, effectively eliminating systematic errors.
4. Simplified operation: Ready-to-use premixed formulation that only needs to be added to the reaction system in proportion; no additional concentration optimization or repeated parameter debugging required.
Product Specifications
Ex/Em: 578/604 nm
Product Components
Component
| 2000 T | | A. 100× Ready-to-use Universal qPCR Reference Dye | 1 mL |
|
Note: 2000 T refers to the total number of 50 μL reaction systems that can be prepared with the kit.
Precautions
1. Shake well before use and avoid repeated freeze-thaw cycles. It is recommended to aliquot Component A into small tubes upon first use and store at -20 ℃.
2. Fluorescent dyes are susceptible to quenching by light exposure; operate in the dark at all times to maintain dye stability.
3. This product is for research use only and must not be stored in ordinary residential premises.
4. For your safety and health, follow the general laboratory safety regulations of your institution.
Instructions for Use
1. Pre-Experiment Preparation
(1) Reagent Preparation
1) Take Component A (100× Ready-to-use Universal qPCR Reference Dye) from the kit and all reagents related to the qPCR/RT-PCR reaction system out of their storage conditions (e.g., -20 ℃ or 4 ℃), and place them on ice or an ice plate.
Note: Protect from light at all times.
2) Instrument Preparation
qPCR instrument
2. Operating Procedures
Prepare the Quantitative PCR/RT-PCR Reaction System
(1) Set up the quantitative PCR/RT-PCR reaction system in accordance with the protocol provided with the PCR reagent kit. For multiple reactions, prepare a master mix first and ensure thorough mixing.
(2) Add Component A (100× Ready-to-use Universal qPCR Reference Dye): Add 0.5 μL of Component A to each 50 μL PCR reaction system.
(3) Mix and centrifuge: Vortex the PCR reaction system to mix well and then perform a brief centrifugation.
Notes:
(1) The volume of the reference dye can be adjusted according to experimental requirements.
(2) No air bubbles or residual liquid on the tube walls shall be present after brief centrifugation.
(3) Protect from light at all times.
| Instrument | 50 µL PCR Reaction | Final Concentration | | Bio-Rad CFS Series | 0 | 0 | | Roche Light Cyler Series | 0 | 0 | | Eppendorf Mastercyler ep realplex Series | 0 | 0 | | Qiagen/Corbelt Rotor-Gene Series | 0 | 0 | | Takara Thermal Cyler Dice Series | 0 | 0 | | ABI PRISM 7000/7300/7700/7900HT and 7900HT Fast system, ABI Step One, ABI Step One Plus Series | 0.5 µL | 1× | | ABI 7500/7500 Fast, ViiA7，QuantStudio, Stratagene Mx3000P/3005P/x4000, MJ Research Series | 0.5 µL | 1× | | Chromo4, Opticon (II), Corbett Rotor Gene 3000 Series | 0.5 µL | 1× |
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ZJ26F0333107
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