The CellAmp Direct TB Green RT-qPCR Kit is designed for performing a simple two-step real-time RT-PCR, without RNA extraction, directly from various animal cells (e.g., adherent cells, floating cells, primary cells, various stem cells, and iPS cells). The kit makes it possible to prepare template samples from cultured cells and complete the analysis of reverse transcription reactions and gene expression in as little as 1.5 hr.
The CellAmp Direct TBGreen RT-qPCR Kit is designed for performing a simple two-step real-time RT-PCR, without RNA extraction, directly from various animal cells (e.g., adherent cells, floating cells, primary cells, various stem cells, and iPS cells). The kit makes it possible to prepare template samples from cultured cells and complete the analysis of reverse transcription reactions and gene expression in as little as 1.5hr.
This product allows the effective elimination of genomic DNA. Therefore, it works well for analyses in which gDNA contamination is an issue, including situations in which the design of a primer pair spanning an exon junction is not possible or in which low-expression genes are to be analyzed. A two-step real-time RT-PCR reagent for TB Green detection is included in the kit, eliminating the need for reverse-transcription and PCR mixes to be prepared separately. This makes gene expression analysis easy to perform.
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请具体点，把联系方式和价位都说一下好吗？
Clontech的非常好, 感觉. 效果不好可能是RNA问题, 或RTase的问题. 因为目前clontech的RTase是Takara的产品，而非原来clontech的产品，可单购。
步骤：1，RNA抽提（同RT）
2，逆转录，使用合成的oligo
3，p内参
4，使用根据已知序列设计的上游引物和提供下游引物做PCR，注意设计上游引物是退火温度与下游引物相差不大。
5，产物切胶回收，TA克隆测序
这个oligo和下游引物是我根据Takara和BD公司的manual自己设计的，经过我的使用效果很好。具体操作参照takara的3－racemanual。你先做3－race，了解熟悉下，成功后再做5－race。5－race要买试剂盒做，较为复杂！
