计算溶液所需的质量、体积或浓度。
为什么选择此级别
，适用于对基线干扰要求严格的色谱和分析工作流程。
储存与运输
避光,-20°C储存,避免反复冻融。超低温运输 。请查阅批次 COA 获取详细规格。
质量文档
SDS、COA、产品数据表及规格说明书均可下载。可通过批号查询获取批次 COA。
文献证明
在色谱分析、有机合成和交叉偶联反应领域已被 0 篇同行评审文献引用。
概述
|
本产品是一步法Real-Time RT-qPCR专用试剂盒。所含的SYBR Green I荧光染料可以与所有的双链DNA相结合，使该产品可以用于多种不同靶序列的检测而不需合成特异性标记探针。使用本产品进行Real Time RT-qPCR反应，逆转录和定量PCR在同一反应体系中进行，反应过程中无需添加试剂，无需打开管盖，避免了污染的同时提高了实验效率。全新高效逆转录酶RNase H活性缺失，减少了逆转录反应中RNA的降解。该酶逆转录效率高，可对少量 RNA模板进行良好的逆转录反应。与RNA亲合性高，能通读GC含量高，二级结构复杂的RNA模板。全新高效热启动酶，在常温下酶的活性被封闭，从而有效避免在常温条件下由引物和模板非特异性结合或引物二聚体而产生的非特异性扩增，极大提高了荧光定量PCR反应的精确性。所包含的缓冲系统使以上两种酶同时发挥最大功效，提高效率。本产品灵敏度高，特异性高，线性范围广，对目的基因定量更准确。ROX染料用于校正定量PCR仪孔与孔之间产生的荧光信号误差，一般用于ABI、Stratagene等公司的Real Time PCR 扩增仪。不同仪器的激发光学系统有所不同，因此ROX染料的浓度必须与相应的荧光定量PCR仪相匹配。
不需要ROX校正的仪器（U665567） : Roche LightCycler 480，Roche LightCyler 96，Bio-rad iCyler iQ，iQ5，CFX96等。
需要High ROX校正的仪器（U665751） ：ABI Prism7000/7300/7700/7900，Eppendorf，ABI Step One/Step One Plus等。
|
注意事项：
1．本试剂盒中的各试剂使用前请上下颠倒轻轻混匀，尽量避免起泡，并经短暂离心后 使用。
2．本产品以RNA为模板进行一步法RT-PCR实验，在操作过程中应避免RNase污染， 建议在专门的区域进行RNA操作，使用专门的仪器和耗材，操作人员带口罩和一次 性手套并经常更换手套，实验相关耗材应用0.1％DEPC(焦碳酸二乙酯)水溶液在37℃ 处理12小时,并高压灭菌30分钟后使用。
3．UltraSYBR One Step RT-qPCR Buffer中含有SYBR Green I 荧光染料，保存本产品 或配制PCR反应液时应避免强光照射。
4．本试剂盒中的各试剂应避免反复冻融，反复冻融可能使产品性能下降。本产品长期 保存可置于-20℃，避光。如果在短期内需要频繁使用，可在2-8℃保存。
5．本试剂盒必须使用特异性引物，引物的选择可根据具体实验来选择，引物设计的好 坏直接影响到RT-PCR反应的结果，设计引物时需考虑GC含量，引物长度，引物位 置，PCR产物的二级结构等因素，建议采用专业的引物设计软件进行设计。
6．本品不能用于探针法荧光定量PCR。
使用方法：
1．将RNA模板、引物、2×UltraSYBR One Step Buffer、UltraSYBR One Step EnzymeMix和RNase-Free Water溶解并置于冰上备用。
2．PCR反应体系：
|
注意：
1）通常引物浓度以0.2 μM可以得到较好结果，可以终浓度0.1-0.5 μM作为设定范围的参 考。扩增效率不高的情况下，可提高引物的浓度；发生非特异性反应时，可降低引物浓度，由此 优化反应体系。
2）不同仪器的激发光学系统有所不同，根据使用荧光定量的仪器选择加入50×Low ROX or 50×High ROX。
3．涡旋震荡混匀，短暂离心，将溶液收集到管底。
4．RT-qPCR反应条件(荧光定量PCR为两步法)，本程序是以ABI 7500荧光定量PCR 仪为例。
|
RT-qPCR反应条件(荧光定量PCR为三步法)：
|
注意：
1）三步法PCR扩增时，退火温度请以 56℃-64℃的范围作为设定参考。
2）融解曲线分析请以所使用的荧光定量PCR仪推荐的程序进行设定，本程序是以ABI
7500荧光定量PCR仪为参照设定。
|
This product is a one-step Real Time RT qPCR specialized reagent kit. The SYBR Green I fluorescent dye contained can bind to all double stranded DNA, making the product suitable for detecting various target sequences without the need for synthesizing specific labeled probes. Using this product for Real Time RT qPCR reaction, reverse transcription and quantitative PCR are performed in the same reaction system, without the need to add reagents or open the tube cap during the reaction process, avoiding contamination and improving experimental efficiency. The loss of RNase H activity, a novel and efficient reverse transcriptase, reduces the degradation of RNA in reverse transcription reactions. This enzyme has high reverse transcription efficiency and can perform good reverse transcription reactions on a small amount of RNA templates. High affinity with RNA, able to read RNA templates with high GC content and complex secondary structures. A new and efficient hot start enzyme, whose activity is blocked at room temperature, effectively avoids non-specific amplification caused by non-specific binding of primers and templates or primer dimers at room temperature, greatly improving the accuracy of fluorescence quantitative PCR reactions. The included buffer system maximizes the effectiveness of both enzymes and improves efficiency. This product has high sensitivity, specificity, wide linear range, and more accurate quantification of target genes. ROX dye is used to correct the fluorescence signal error between wells in quantitative PCR instruments, and is generally used in Real Time PCR amplification instruments from companies such as ABI and Stratagene. The excitation optical systems of different instruments vary, so the concentration of ROX dye must be matched with the corresponding fluorescence quantitative PCR instrument.
Instruments that do not require ROX calibration( U665567 ): Roche LightCycle 480, Roche LightCyler 96, Bio rad iCyler iQ, iQ5, CFX96, etc.
Instruments that require High ROX calibration include( U665751）：ABI Prism7000/7300/7700/7900, Eppendorf, ABI Step One/Step One Plus, etc.
|
Notes:
1. Before use, please gently mix the reagents in this reagent kit upside down to avoid foaming, and use after brief centrifugation.
2. This product uses RNA as a template for one-step RT-PCR experiments. During the operation, RNase contamination should be avoided. It is recommended to perform RNA operations in a dedicated area, using specialized instruments and consumables. Operators should wear masks and disposable gloves and frequently change gloves. The experimental consumables should be treated with 0.1% DEPC (diethyl pyrocarbonate) aqueous solution at 37 ℃ for 12 hours and sterilized under high pressure for 30 minutes before use.
3. UltraSYBR One Step RT qPCR Buffer contains SYBR Green I fluorescent dye. When storing this product or preparing PCR reaction solution, avoid strong light irradiation.
4. Each reagent in this kit should avoid repeated freeze-thaw cycles, as repeated freeze-thaw cycles may lead to a decrease in product performance. This product can be stored for a long time at -20 ℃, away from light. If frequent use is required in the short term, it can be stored at 2-8 ℃.
5. This reagent kit must use specific primers, and the selection of primers can be based on specific experiments. The quality of primer design directly affects the results of RT-PCR reactions. When designing primers, factors such as GC content, primer length, primer position, and the secondary structure of PCR products need to be considered. It is recommended to use professional primer design software for design.
6. This product cannot be used for probe based fluorescence quantitative PCR.
Usage:
1. Dissolve the RNA template, primers, 2x UltraSYBR One Step Buffer, UltraSYBR One Step EnzymeMix, and RNase Free Water and place them on ice for later use.
2. PCR reaction system:
|
Attention:
1) Typically, the primer concentration is 0.2 μ M can obtain good results, with a final concentration of 0.1-0.5 μ M serves as a reference for setting the range. In the case of low amplification efficiency, the concentration of primers can be increased; When non-specific reactions occur, the primer concentration can be reduced to optimize the reaction system.
2) The excitation optical systems of different instruments vary, and depending on the instrument used for fluorescence quantification, 50 x Low ROX or 50 x High ROX can be added.
3. Vortex and shake well, centrifuge briefly, and collect the solution to the bottom of the tube.
4. RT qPCR reaction conditions (fluorescence quantitative PCR is a two-step method). This program takes the ABI 7500 fluorescence quantitative PCR instrument as an example.
|
RT qPCR reaction conditions (fluorescence quantitative PCR is a three-step method):
|
Attention:
1) When using the three-step PCR amplification method, the annealing temperature should be set within the range of 56 ℃ -64 ℃ as a reference.
2) Please set the fusion curve analysis according to the recommended program of the fluorescence quantitative PCR instrument used. This program is based on the ABI 7500 fluorescence quantitative PCR instrument as a reference setting.
规格
别名
UltraSYBR 一步法 RT-qPCR 试剂盒
英文名称
UltraSYBR One Step RT-qPCR Kit
储存条件
避光,-20°C储存,避免反复冻融
运输条件
超低温运输
产品线
生物产品
名称和识别符
分子类型
试剂盒
技术文档
高级数据
质检证书（CoA,COO,BSE/TSE 和分析图谱)
C of A & Other Certificates(BSE/TSE, COO):
输入批号以搜索分析图谱:
溶液计算器
常见问题
本产品应如何储存？
请于?20 °C、避光条件下保存。避免反复冻融。建议按单次用量分装保存。
本产品如何运输？
本产品采用保温箱配冰袋运输。收货后请立即拆包，并按上述储存条件保存。
如何获取批次专用COA？
每个订单都会随附分析证书（COA），您也可以从账户订单历史中下载。如需订购前的COA，请联系您的客户代表，或使用PDP上的"申请COA"按钮——我们将提供代表性批次的COA。
产品仅提供给经过验证的企业、机构和合格的专业人员用于研发。不适用于人类、动物、诊断或治疗。
Copyright © 2023至今 上海阿拉丁生化科技股份有限公司 保留所有权利。
危险品化学品经营许可证（带存储）