谁做过PCR?We carried out an anchored RT-PCR and cloned the low levels of Foxp3 mRNA in the breast tissues.anchored RT-PCR

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谁做过PCR?We carried out an anchored RT-PCR and cloned the low levels of Foxp3 mRNA in the breast tissues.anchored RT-PCR
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谁做过PCR?We carried out an anchored RT-PCR and cloned the low levels of Foxp3 mRNA in the breast tissues.anchored RT-PCR
谁做过PCR?
We carried out an anchored RT-PCR and cloned the low levels of Foxp3 mRNA in the breast tissues.
anchored RT-PCR

谁做过PCR?We carried out an anchored RT-PCR and cloned the low levels of Foxp3 mRNA in the breast tissues.anchored RT-PCR
anchored RT-PCR
锚定RT-PCR
锚定PCR
一般采用的PCR技术必须首先知道欲扩增DNA或RNA片段两侧的序列,而大多数情况下对某些序列本身或其旁侧序列并不清楚,这就限制了PCR技术的广泛应用.“锚定PCR(anchored PCR)”可以克服序列未知或未全知带来的障碍.该法适用于mRNA的扩增,而不适用于基因组DNA的扩增.首先分离细胞总RNA或mRNA,在反转录酶作用下合成cDNA,然后在DNA末端转移酶作用下,在cDNA3’端加上poly(dG)尾巴,最后用特异性的3’端引物和一种锚定引物poly(dC)一起做PCR扩增.为保证扩增特异性,建议此段寡聚(dC)在十二聚体以上,其5’端可带上某些限制酶序列或其他序列信息.用此方法使带同源多聚物尾巴的cDNA序列得以扩增.
锚定RT-PCR扩增的具体操作步骤
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