RT2 PreAMP Pathway Primer Mixes

用于RT² Profiler PCR Arrays分析之前扩增cDNA模板

S_1084_5_GEN_V2
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RT2 PreAMP Pathway Primer Mix

Cat. No. / ID:  330241

RT2 Nano PreAMP Primer Mix
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RT2 PreAMP Pathway Primer Mixes 旨在用于分子生物学应用。该产品不能用于疾病诊断、预防和治疗。
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Features

  • 适用于人类、小鼠和大鼠的引物混合液
  • 最少的手动操作时间
  • 对少量样本也可灵敏检测

Product Details

RT² PreAMP cDNA Synthesis Kit和RT² PreAMP Pathway Primer Mixes是创新技术,能够用低至1 ng的总RNA进行基因表达分析。专有技术的扩增过程增加了用于PCR芯片分析的cDNA的量。起始样本包括穿刺活检、激光捕获显微解剖样本、干细胞团或胚胎体及流式细胞分选仪(FACS)制备的细胞。专门设计的引物混合液适用于人类、小鼠或大鼠的Custom RT² Profiler PCR Arrays。

Performance

阳性信号率更高,无偏差扩增获得无偏向性的结果

使用RT² PreAMP Pathway Primer Mix进行预扩增之后,可检测的基因更多(参见" Increased positive call rate")。

扩增过程无偏向性,回归分析评估发现,预扩增的与未扩增的cDNA之间能产生高度可比较的ΔCT值(参见" Unbiased amplification process")。

在预扩增与未扩增样本之间,基因表达倍数变化有高度相关性(参见" Faithfully amplified biology")。

See figures

Principle

RT² PreAMP技术利用多重串联PCR以最小的偏差预扩增基因特异性cDNA。根据逆转录本,RT² PreAMP Pathway Primer Mix用于扩增模板,能够在4个Custom RT² Profiler PCR Arrays上进行基因表达分析(参见" Simple template amplification and gene-expression analysis")。
See figures

Procedure

首先使用RT² PreAMP cDNA Synthesis Kit同时反转录最多12 种不同的RNA样本为cDNA。然后将cDNA进行预扩增用于特定的基因列表。每一个反转录反应用4种RT² PreAMP Pathway Primer Mixes进行扩增,能够在4个特定PCR 芯片上进行基因表达分析。RT²  PreAMP cDNA Synthesis Kit中含有Side Reaction Reducer,可去除预扩增所产生的引物残留,能在RT² Profiler PCR Arrays进行准确检测。把预扩增的模板与仪器特异性、即用型RT² SYBR® Green Mastermix混合, 便完成PCR芯片过程。

Applications

使用RT2 PreAMP cDNA Synthesis Kit及RT2 PreAMP Pathway Primer Mixes生成的cDNA模板可即用于RT2 Profiler PCR Arrays进行的基因表达谱的分析。

Supporting data and figures

Resources

安全数据表 (1)
Download Safety Data Sheets for QIAGEN product components.
产品介绍与指南 (1)
Simultaneously profile mRNA, miRNA and lncRNA using a simple, complete workflow
Safety Data Sheets (1)
Certificates of Analysis (1)
Brochures & Guides (1)
Simultaneously profile mRNA, miRNA and lncRNA using a simple, complete workflow

FAQ

Is it good to pool multiple RNA replicates to detect expression changes that are consistently reproducible?
With the additional RT2 PreAMP methodology, only 1 ng of RNA is now needed for PCR Array analysis. Pooling RNA from different sources should only be done when there is not enough sample. We recommend running biological replicates.
FAQ ID -2663
How do I create a workspace that is free of DNA contamination, prior to carrying out a qPCR experiment?

Any DNA contamination will artificially inflate the SYBR Green signal, yielding skewed gene expression profiles and false-positive signals. The most common source of DNA contamination is from PCR products generated during previous experiments. Such contamination is most often due to the improper disposal of tubes, tips, and gels that previously came into contact with PCR products. Additionally, PCR products may also contaminate pipettors, racks, work pads, and commonly used reagents such as water and buffers. To minimize the risk of contaminating your experiment with extraneous DNA, the following steps should be taken:

 

  • Remove a single aliquot of water from your PCR-grade stock, sufficient to complete the experiment. This minimizes the number of times that the stock container is opened, thereby minimizing contamination risks.
  • Use only fresh PCR-grade reagents and disposable labware.
  • Treat any labware (tubes, tips, and tip boxes) used in PCR with 10% bleach, before discarding.
  • Maintain a dedicated workspace for PCR setup (perhaps a PCR-only hood), away from areas of the lab where post-PCR work is done, such as running gels, enzyme digestions, and cloning.
  • Change the lab bench pads/papers often and decontaminate lab benches and labware (racks, pipettors, etc.) before each use by washing with 10% bleach, and/or exposing to UV light for at least 10 minutes. This serves to degrade and/or inactivate contaminating DNA.
  • Before, during, and after the experiment, minimize the opening and closing of any tubes or plates used during the experiment.  
FAQ ID -2654