Superior stability after reaction setup at room temperature
扩增GC含量高的模板
Amplification of long amplicons
QIAGEN PCR缓冲液中的NH4+和K+促进引物特异性结合
可选的加样变色指示剂
卓越的灵敏性
超群的特异性
OneStep Ahead RT-PCR procedure
HeLa total RNA (10 and 1 pg) was used as a template for amplification of ACTB in triplicates, according to suppliers’ instructions. Reactions were either set up on ice or left at room temperature for the times indicated before analysis on a 2% agarose gel. Distinct, gene-specific bands are observed with the QIAGEN OneStep
Ahead
RT-PCR Kit even after a 2 h incubation at room temperature before cycling (blue arrow), whereas reactions performed with kits from competitors deteriorate as time progresses. Gene-specific bands appear significantly weaker, if present at all, while primer-dimers (red arrows) become more prominent.
将HeLa总RNA（100 ng）作为模板，扩增GC含量达67.1%的TNFRI (581 bp)。反应平行进行三次，绿色箭头表示特异性产物。在QIAGEN产品的反应中使用了Q-Solution。
HeLa total RNA (100 and 10 ng) was used as a template for amplification of RANBP2 (3 kbp).
钾离子结合到DNA骨架上的磷酸基团，可稳定引物结合到模板。在热循环条件下，NH
4+
同时以铵离子和氨的形式存在，会与碱基对之间的氢键相互作用，使错配碱基之间的氢键不稳定。上述两种离子的作用使得在宽泛的温度范围内保持高的特异性结合。
该试剂盒提供了一种特殊的加样变色指示剂。预混液变色指示剂为橙色的惰性染料，可加入预混液中。模板变色指示剂为蓝色的惰性染料，可加入模板中。将模板加入预混液后，反应体系颜色将会有橙色变为绿色，从而可便利的观察移液的进展情况。此外，这两种染料都可在凝胶电泳时作为示踪染料。在1%的琼脂糖凝胶上，橙色染料可指示50 bp条带的位置，而蓝色为4000 bp。
将图示量的HeLa总RNA（单位为pg）作为模板，扩增GAPDH (831 bp)和ACTB (295 bp)，反应平行进行两次。绿色箭头表示特异性产物，红色箭头为引物二聚体。后续分析在QIAxcel上进行。
将图示量的HeLa总RNA（单位为pg）作为模板，扩增EIF2B4 (107 bp)，反应平行进行两次。绿色箭头表示特异性产物，红色箭头为引物二聚体，粉色箭头表示非特异性扩增产物。后续分析在QIAxcel上进行。
The QIAGEN OneStep
Ahead
RT-PCR Kit allows fast and easy RT-PCR setup whatever your application - virus detection, molecular diagnostics research or gene expression - just mix all components together in one tube and start your thermal-cycler program. The reaction mixture contains all of the reagents required for both reverse transcription and PCR; nothing needs to be added once the reaction has been started. As a visual pipetting control, you can add the optional orange Master Mix Tracer and blue Template Tracer.