这道题怎么做？求帮忙 感谢老师主题班会ppt万分

来源：蜘蛛抓取(WebSpider) 时间：2017-09-11 13:34 标签： 英文感谢邮件主题
跪求大神帮忙做一下这道数学题，万分感谢~【中科大研究生吧】_百度贴吧
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跪求大神帮忙做一下这道数学题，万分感谢~收藏
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登录百度帐号推荐应用Reviewer提的两个问题，求如何回答，如何做？万分感谢 - 实验交流 - 生物秀
标题: Reviewer提的两个问题，求如何回答，如何做？万分感谢
摘要: [Reviewer提的两个问题，求如何回答，如何做？万分感谢] Reviewer提的两个问题，本人对这个问题有点拿不准，求热心战友帮忙，如何回答？，如何做？万分感谢Reviewer
2: The authors did address most concerns of the reviewer
However, there is one technical 关键词:[内含子 引物 标准曲线 阳性对照 反转录酶 条带 对照实验]……
Reviewer提的两个问题，本人对这个问题有点拿不准，求热心战友帮忙，如何回答？，如何做？万分感谢
Reviewer #2: The authors did address most concerns of the reviewer. However, there is one technical issue which has to be clarified.
1. In the MM section, the authors state that &To ensure that the samples were free of genomic DNA, RNA samples were subjected to a reverse transcription in the absence of the reverse transcriptase. Our results didn"t find any genomic DNA contamination.& The authors are correct to assume that if there is any contamination with genomic DNA, then there should be amplification in the subsequent PCR. HOWEVER, in case the primer is an exon-intron spanning oligonucleotide and the amplified intron is too long, than this genomic contamination band will not be visible in a 1% agarose gel. I strongly suggest performing another control experiment with contaminated RNA to ensure that the assay performed is sufficient to detect genomic DNA contamination.
3. Which strategy was followed to evaluate real-time PCR experiments?回复第一个说得很明白吧?让你补试验你就老实补吧.虽然似乎有点挑剔:D
我理解就是让你加个阳性对照,把RNA里面故意加入一点genomic DNA.然后用你的引物应该可以在胶上看到污染.回复1. 审稿者认为你的实验的对照有问题。你的对照是不加，如果有条带的话，就是DNA污染，但是，由于DNA含有内含子，如果内含子长度过长而且你的引物是跨内含子设计的，那么你依旧不能扩出条带了，但是这种情况下依旧会有DNA污染，所以审稿人让你再设计其他的对照实验，以避免这样的问题。或者，如果你有证据证明你的引物不是跨内含子设计的，那么只向审稿人说明这个问题，并在方法里也相应加上说明就行了。
3.是问评估real-time PCR的方法。也就是你用什么方法来在证明你用这这个real-time PCR体系是有效的。一般来说，都是通过预实验绘制标准曲线来评估real-time PCR体系的优劣的。 每对引物在正式试验之前都需要做一个标准曲线，看获得的标准曲线的斜率是否符合标准。回复
第一个说得很明白吧?让你补试验你就老实补吧.虽然似乎有点挑剔:D
我理解就是让你加个阳性对照,把RNA里面故意加入一点genomic DNA.然后用你的引物应该可以在胶上看到污染. 谢谢回复
1. 审稿者认为你的实验的对照有问题。你的对照是不加，如果有条带的话，就是污染，但是，由于含有内含子，如果内含子长度过长而且你的引物是跨内含子设计的，那么你依旧不能扩出条带了，但是这种情况 ... 非常感谢
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&&Reviewer提的两个问题，求如何回答，如何做？万分感谢
Reviewer提的两个问题，求如何回答，如何做？万分感谢
Reviewer提的两个问题，本人对这个问题有点拿不准，求热心战友帮忙，如何回答？，如何做？万分感谢
Reviewer #2: The authors did address most concerns of the reviewer. However, there is one technical issue which has to be clarified.
1. In the MM section, the authors state that &To ensure that the samples were free of genomic DNA, RNA samples were subjected to a reverse transcription in the absence of the reverse transcriptase. Our results didn't find any genomic DNA contamination.& The authors are correct to assume that if there is any contamination with genomic DNA, then there should be amplification in the subsequent PCR. HOWEVER, in case the primer is an exon-intron spanning oligonucleotide and the amplified intron is too long, than this genomic contamination band will not be visible in a 1% agarose gel. I strongly suggest performing another control experiment with contaminated RNA to ensure that the assay performed is sufficient to detect genomic DNA contamination.
3. Which strategy was followed to evaluate real-time PCR experiments?
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