※ 引述《weisont (我的腰好酸)》之铭言： : 这应该是两回事吧 : PCR是PCR : CLONE是CLONE : 两者并不是有相同功能而可以互相取代的 : PCR是将目标基因放大 : CLONE的目的一部分是保存目标基因 : 一部分则是做INVIVO的实验 : 如果不做CLONE : 那要怎麽来表现这段基因勒?? 喔 原来我刚刚误会了~~ [因为我太累 懒的看英文...||] 原来呢..作clone的目的 是想得到这段基因表现出来的rRNA 然後比较rRNA的特殊二级结构 就可以确定PCR的过程中有没有出状况.. 但是因为现在PCR的技术越来越好 所以已经不用透过这个步骤来confirm了... 嗯..应该是吧..[我还是没有看下面那篇文章...||] 所以 有问题再提出来吧XD --- 以下是资料的原文.. In summary, there appears to be no reason for more than the usual concern, because of Taq polymerase fidelity of replication, about sequence accuracy in this method. Several positions that had previously been scored as unknown or ambiguous are now determinable by taking advantage of bidirectional gene sequencing and the absence of artifacts due to rRNA secondary structure and posttranscriptionally modified nucleotides. As always, a cautious approach should be taken by checking any sequence anomalies and confirming known secondary structural constraints on sequences. Although we have spent considerable time optimizing and testing the direct sequencing of PCR-amplified rDNA, we highly recommend cloning the fragments if a near-perfect sequence is desired. ........ The amplification by PCR of a taxonomically diverse collection of eubacterial 16S rDNA genes is possible with a small number of primers. These products can readily be cloned for sequencing or they can be sequenced directly. -- I'm just a small potato. --

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