
The Yeast Processing Reagent (for total RNA preparation) is a pretreatment reagent that may be used in combination with RNApurificationreagents toenable RNA isolation from yeast. Pretreatment of yeast cells with this product comprises two steps: 1) washing and recovery of yeast cells by centrifugation, and 2) yeast cell wall disruption via a Yeast Processing Enzyme Solution (containing a cell wall degradation enzyme) and a Yeast Processing Buffer.
The Yeast Processing Reagent (for total RNA preparation) is a pretreatment reagent that may be used in combination with RNApurificationreagents toenable RNA isolation from yeast. Pretreatment of yeast cells with this product comprises two steps: 1) washing and recovery of yeast cells by centrifugation, and 2) yeast cell wall disruption via a Yeast Processing Enzyme Solution (containing a cell wall degradation enzyme) and a Yeast Processing Buffer. Becauseyeast cell walldisruption occurs throughenzymatic activity, the time-consuming cell-lysis protocols using glass beads and liquid nitrogen required in conventional methods are unnecessary.
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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!

