CellsDirect™ 一步法 qRT-PCR 试剂盒
CellsDirect™ 一步法 qRT-PCR 试剂盒
Invitrogen™

CellsDirect™ 一步法 qRT-PCR 试剂盒

CellsDirect™ 一步法 qRT-PCR 试剂盒可直接从细胞中获得高灵敏度和特异性的实时 qPCR 结果,无需 RNA 纯化步骤。CellsDirect™ 一步法了解更多信息
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货号反应次数
11753100100 次反应
11753500500 次反应
货号 11753100
价格(CNY)
8,463.00
Each
添加至购物车
反应次数:
100 次反应
价格(CNY)
8,463.00
Each
添加至购物车
CellsDirect™ 一步法 qRT-PCR 试剂盒可直接从细胞中获得高灵敏度和特异性的实时 qPCR 结果,无需 RNA 纯化步骤。

CellsDirect™ 一步法 qRT-PCR 试剂盒的特点包括:

高效—节省大量时间、降低成本、提高通量
便利—酶混合物包括 SuperScript™ III 逆转录酶和 Platinum™ Taq DNA 聚合酶
灵敏度—没有单独的 RNA 分离步骤,可较大程度减少样品损失,提高灵敏度
检测范围宽—从单个细胞至 10,000 个细胞范围的样品检测靶标
多功能性—与激光捕获显微切割 (LCM) 收集的多种细胞系和样品兼容

简化工作流程
在传统的实时荧光定量 RT-PCR (qRT-PCR) 中,首先通过耗时的程序从细胞分离 RNA,该程序可导致材料损失。使用 CellsDirect™ 一步法 qRT-PCR 试剂盒,可以裂解细胞并直接将全部裂解物加入一步法 qRT-PCR 反应中,实现较少的处理和样品损失。

非常适合 qRT-PCR 应用
CellsDirect™ 技术经过验证,可用于涉及有限样品的基因表达研究。CellsDirect™ 一步法 qRT-PCR 试剂盒通过消除昂贵、耗时的 RNA 纯化步骤对于高通量应用特别有价值(参见图)。CellsDirect™ 一步法 qRT-PCR 试剂盒也可用于检测基因组 DNA 靶标。

CellsDirect™ 一步法 qRT-PCR 试剂盒可与水解探针(例如,TaqMan™ 或 LUX™ 荧光引物)配合使用。

注:该试剂盒包含一管单独的 ROX™ 参比染料。另外,还提供 2X 反应混合物中包含 ROX™ 参比染料的等效试剂盒 (11754-500)。
仅供科研使用。不可用于诊断程序。
规格
适用于(设备)7500 Fast 系统、7500 系统、BioRad iQ5、BioRad MyiQ、Corbett RotorGene、MJ Opticon、Stratagene Mx3000P、Stratagene Mx3005P、Stratagene Mx4000、BioRad iCycler iQ、Cepheid SmartCycler、MJ Chromo4、QuantStudio 3 和 5 系统、QuantStudio 6 和 7 Pro 系统、QuantStudio 7 Flex 系统、QuantStudio 12k Flex 系统、QuantStudio 6 Flex 系统
反应次数100 次反应
产品线CellsDirect、Platinum、SuperScript
数量100次反应
样品类型细胞
最大浓度2X
检测方法引物-探针
适用于(应用)RT-PCR
反应速度快速,标准
Unit SizeEach
内容与储存
•10 mL 重悬缓冲液
•1 mL 裂解增强剂
•500 µL DNase I(扩增级)(1 U/µL)
•160 µL 10X DNase I 缓冲液
•400 µL 25 mM EDTA
•100 µL SuperScript ™III RT/Platinum™Taq 混合物
•2 × 1.25 mL 2X 反应混合物
• 1 mL 50 mM MgSO4
•2 mL DEPC 处理水
•10 µL HeLa 总 RNA (10 ng/µL)
•100 µL ROX™ 参比染料(在 -20°C 下于暗处储存)
该试剂盒置于干冰上运输,并应将其储存在 -20°C 下

常见问题解答 (FAQ)

What can I do to improve the sensitivity of my qPCR assay?

If you are targeting a low-abundance gene, you may have trouble getting Ct values in a good, reliable range (Ct > 32). To increase the sensitivity of the assay, you may want to consider the following:

- Increase the amount of RNA input into your reverse transcription reaction, if possible
- Increase the amount of cDNA in your qPCR reaction (20% by volume max)
- Try a different reverse transcription kit, such as our SuperScript VILO Master Mix, for the highest cDNA yield possible
- Consider trying a one-step or Cells-to-CT type workflow (depending on your sample type)

How do I set the baseline for my qPCR experiment?

Most times your instrument software can automatically set a proper baseline for your data. Check out our short video, Understanding Baselines, for more information on how to set them (https://www.youtube.com/watch?feature=player_embedded&v=5BjFAJHW-bE).

How do I set the threshold for my qPCR experiment?

In most cases your instrument software can automatically set a proper threshold for your data. Check out our short video, Understanding Thresholds, for more information on how to set them (https://www.youtube.com/watch?feature=player_embedded&v=H_xsuRQIM9M).

I am not getting any amplification with my TaqMan Assay or SYBR Green primer set. What is causing this?

There could be several reasons for no amplification from an assay or primer set. Please see these examples and suggested solutions in our Real-Time Troubleshooting Tool (https://www.thermofisher.com/us/en/home/life-science/pcr/real-time-pcr/qpcr-education/real-time-pcr-troubleshooting-tool/gene-expression-quantitation-troubleshooting/no-amplification.html) for more details.

I am getting amplification in my no-template control (NTC) wells in my qPCR experiment. What is causing this?

There could be several reasons for amplification in a NTC well. Please see these examples and suggested solutions in our Real-Time Troubleshooting Tool (https://www.thermofisher.com/us/en/home/life-science/pcr/real-time-pcr/qpcr-education/real-time-pcr-troubleshooting-tool/gene-expression-quantitation-troubleshooting/amplification-no-template-control.html) for more details.

引用和文献 (1)

引用和文献
Abstract
Comparison of reverse transcription-quantitative polymerase chain reaction methods and platforms for single cell gene expression analysis.
Authors:Fox BC, Devonshire AS, Baradez MO, Marshall D, Foy CA,
Journal:Anal Biochem
PubMed ID:22617801
Single cell gene expression analysis can provide insights into development and disease progression by profiling individual cellular responses as opposed to reporting the global average of a population. Reverse transcription-quantitative polymerase chain reaction (RT-qPCR) is the  ... More