Zero Blunt™ TOPO PCR Cloning Kit for Subcloning - FAQs

查看更多产品信息 Zero Blunt™ TOPO PCR Cloning Kit for Subcloning - FAQs (K283020)

2 个常见问题解答

TOPO TA 克隆载体和 Zero Blunt 克隆载体能否用于体外转录以生成探针？它们可以用来做体外翻译吗？

1. 是的，TOPO TA Cloning 和Zero Blunt cloning vectors可以用于体外转录。这些载体含有的T7、SP6 或T3启动子是活跃的噬菌体启动子，相应的聚合酶将从这些启动子转录RNA。

2. 这些载体通常不建议用于体外翻译，因为这些载体的启动子下游（3’端）大多含有一个或多个翻译起始信号（ATG）。ATG在多克隆位点之内，这可能会干扰在插入片段中开放阅读框的翻译。

Can TOPO TA cloning vectors and Zero Blunt cloning vectors be used for in vitro transcription to generate probes? Can they be used to do in vitro translation?

Yes, the TOPO TA cloning and Zero Blunt cloning vectors can be used for in vitro transcription. The T7, SP6, and T3 promoters contained in these vectors are active phage promoters, and the corresponding polymerases will transcribe RNA from these promoters.
These vectors are not generally recommended for in vitro translation because most of these vectors contain one or more translational initiation signals (ATG) downstream (3’) of the promoters contained in them, within the multiple cloning sites, which may interfere with attempts to translate an open reading frame in the insert.