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lhazrael

木虫 (正式写手)

应助: 7 (幼儿园)
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注册: 2009-02-27
性别: MM
专业: 微生物遗传育种学

[求助] 生物类的请帮忙看下审稿意见，急！！

急着毕业所以投的是biotechnology letters，不知道是为什么就给了一个审稿人的意见，下面是审稿建议：
- Using RT-PCR rather than qRT-PCR is risky and may lead to misleading results. It is unclear to me whether the gel presented in fig 1 is a representative of several independent experiments or just a one-shot test. I would strongly urge the authors to use qPCR to determine the level of RNAi in teh transpormants. The number of replicats used should be indicated for all of the data presented (not just fig 1). （实验结果的确是一次实验得到的one-shot test）
- I have the same question for the growth data. Are the pictures shown the representative of several biological replicats and if so, was there any variation in the growth for the transformants, or is this just a single experiment.
- The absence of a correlation between the effect on growth and on conidiation requires more discussion (assuming this is reproducable) and at least an hypothesis. Based on the current text the authors suggest that donwregulation of the chitin synthase results in reduced growth and conidiation, but based on the current data these effects are not linked. An explanation or hypothesis is needed.
- The correlation between expression and penicillin production also does not fit. E.g. transformants 10, 11 and 15 have similar expression as the original strain but significantly reduced production. This could again be due to an artefact of the RT-reaction, strengthening the need for qPCR.

1.实验室做分子很少，qPCR更没做过，感觉做rt-pcr已经很崩溃了，再摸索估计耗很长时间，看的相关文章也有直接做RT-PCR的，能不能绕过不做qPCR啊？
2.第一个问题中的The number of replicats是不是指实验重复次数？
3.第二个问题，我的实验数据中都有error bar的，除了菌落和菌丝照片，他为什么还这么问呢？
4.关于第三个问题，我做的实验也是参照国外的文献做的，别人的结果就这样啊，也没细讲，再说中间的关系肯定复杂的很啊，怎么回答？
5.第四个问题，影响产量的因素很多啊，我文章中有大体解释过，产量的下降跟发酵过程中的传质传氧结球有很大关系的，但因为是letters，图片数文章篇幅都有限制，不可能细讲的啊

希望高手帮忙啊，急！！

[ Last edited by lhazrael on 2012-10-7 at 14:16 ]

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