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RT2 PreAMP Pathway Primer Mixes

用于RT² Profiler PCR Arrays分析之前扩增cDNA模板

  • 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

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。

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产品 Product no. 货号 目录价:
 
 
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RT2 PreAMP Pathway Primer Mixes适用于分子生物学应用。该产品不适用于疾病的诊断、预防或治疗。


0
Increased positive call rate.
First strand cDNA was synthesized from 40 ng mouse total RNA with (light blue bars) or without (dark blue bars) preamplification with RT² PreAMP PCR Mastermix and RT² PreAMP Pathway Primer Mix. The unamplified and amplified samples were then analyzed on the Mouse Inflammatory Cytokines & Receptors RT² Profiler PCR Array and the threshold CT were obtained. Genes with CT ≥35 were considered to be "absent". In the unamplified sample, 75% of the genes analyzed were present. The positive call rate was increased to 100% in the amplified sample. The 22 genes shown were called "absent" in the unamplified sample and were detectable in the preamplified sample.
1
Simple template amplification and gene-expression analysis.
2
Unbiased amplification process.
First strand cDNA was synthesized from 5 ng of human liver tumor RNA. One-quarter of the first strand cDNA was then used for preamplification with RT² PreAMP PCR Master Mix plus Human Cancer PathwayFinder RT² PreAMP Pathway Primer Mix. Unamplified cDNA synthesized from 500 ng of the same sample was used as a control. Preamplified and unamplified cDNA samples were then analyzed on the Human Cancer PathwayFinder RT² Profiler PCR Array and the DCT values were obtained. Concordance between preamplified and unamplified samples was evaluated by regression analysis. Data points with CT ≥35 were considered to be “absent” genes and were excluded from analysis. The lines show a linear regression fit with the R2 and slope indicated in the graph.
3
Faithfully amplified biology.
First strand cDNA was synthesized from 1 ng human liver tumor RNA or universal RNA. One-quarter of the first strand cDNA was then used for preamplification with RT² PreAMP PCR Master Mix plus Human Cancer PathwayFinder RT² PreAMP Pathway Primer Mix. Unamplified cDNA synthesized from 500 ng of each corresponding RNA sample was used as a control. Preamplified and unamplified cDNA samples were analyzed on the Human Cancer PathwayFinder RT² Profiler PCR Array and the fold change in gene expression between liver tumor and universal RNA for each gene was obtained using the ΔΔCT method.
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")。

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")。
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。

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试剂盒操作手册 (1)
For synthesis and preamplification of cDNA from small RNA samples and RNA from formalin-fixed, paraffin-embedded samples
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