采用大引物方法，利用质粒多克隆位点两侧的普通测序引物作为旁侧引物，在单个PCR管内，经2个步骤共34个循环进行定点突变。

Using normal sequencing primers adjacent to multiple clone sites of plasmid as flanking primers, a novel megaprimer method was performed in one tube with two steps and 34 cycles.

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The rest of this, this sort of beige part of the molecule here is called the polylinker part.

剩下这部分,这部分淡棕色的分子叫做多克隆位点

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How we do that will be clear in a few minutes, but this polylinker as is described down here is a site where you can clone in genes.

马上你们就清楚这该怎么做了,这里所说的多克隆位点,就是你能把基因片段插进来的位点

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We're going to have DNA that we would like to make lots of copies of, or we're going to have DNA that we would like to get expressed in a cell, and we're going to put it in this region that's called the polylinker.

我们将那些需要得到很多拷贝数的DNA,或者需要在细胞中表达的DNA,整合到这个叫做多克隆位点的区域中

耶鲁公开课 - 生物医学工程探索课程节选