【分享】高手手把手教你如何选择realtime qPCR的数据分析方法
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一般的数据分析方法的文献光看中文就够头大,但是这个英文文献很通俗易懂,很适合我们初学者。
文中列举了几个实际的例子,可以方便大家参考。
Example 1
The mean CT of the HOXD10 gene in treated and untreated samples was 24.6 and 27.5, respectively. The mean CT of the 18S rRNA in the treated and untreated samples was 9.9 and 9.8, respectively. What is the fold change in expression of the HOXD10 gene due to treatment?
Example 2
The mean CT of the HOXD10 gene in treated and untreated samples was 26.5 and 24.9, respectively. The mean CT of the 18S rRNA internal control in the treated and untreated samples was 9.7 and 9.9, respectively. What is the fold change in expression of the HOXD10 gene due to treatment?
Example 3
The gene expression is to be compared in cell cultures that are treated with a hypothetical drug to those that are untreated. The mean CT from replicate runs of an internal control gene are 27.2, 27.0 and 27.4 (treated samples) and 26.2, 26.3 and 26.0 (untreated samples). What is the fold change in expression of the internal control in the treated versus the untreated samples? Does this gene serve as a useful internal control in this experiment?
Example 4
The expression of the BCL-2 and 18S rRNA genes were quanti?ed in a group of 20 HER2 positive breast cancer patients and in 20 HER2 negative breast cancer patients. What is the mean ± s.d. for the HER2 expression in each group? What is the difference in the Bcl-2 expression in the HER2 positive compared with the HER2 negative patients?
Example 5
As another example, let us assume that three mice were treated with an experimental drug and three mice were untreated. cDNA was synthesized from the livers of each mouse and triplicate PCRs were performed on each sample of cDNA. The mean CT (treated mice) from the triplicate PCRs for the target and internal control gene are 27.2, 28.0, 27.8 (target gene) and 24.2, 24.7 and 24.9 (internal control gene). The mean CT (untreated mice) from the triplicate PCRs for the target and internal control gene are 22.8, 23.0, 22.4 (target gene) and 24.8, 25.0 and 24.7 (internal control gene). What are the mean ± s.d. and the coef?cient of variation (CV) for the relative amount of target gene in both groups of mice? What is the difference in expression of the target gene in the liver of the treated mice compared to the untreated mice?
文中列举了几个实际的例子,可以方便大家参考。
Example 1
The mean CT of the HOXD10 gene in treated and untreated samples was 24.6 and 27.5, respectively. The mean CT of the 18S rRNA in the treated and untreated samples was 9.9 and 9.8, respectively. What is the fold change in expression of the HOXD10 gene due to treatment?
Example 2
The mean CT of the HOXD10 gene in treated and untreated samples was 26.5 and 24.9, respectively. The mean CT of the 18S rRNA internal control in the treated and untreated samples was 9.7 and 9.9, respectively. What is the fold change in expression of the HOXD10 gene due to treatment?
Example 3
The gene expression is to be compared in cell cultures that are treated with a hypothetical drug to those that are untreated. The mean CT from replicate runs of an internal control gene are 27.2, 27.0 and 27.4 (treated samples) and 26.2, 26.3 and 26.0 (untreated samples). What is the fold change in expression of the internal control in the treated versus the untreated samples? Does this gene serve as a useful internal control in this experiment?
Example 4
The expression of the BCL-2 and 18S rRNA genes were quanti?ed in a group of 20 HER2 positive breast cancer patients and in 20 HER2 negative breast cancer patients. What is the mean ± s.d. for the HER2 expression in each group? What is the difference in the Bcl-2 expression in the HER2 positive compared with the HER2 negative patients?
Example 5
As another example, let us assume that three mice were treated with an experimental drug and three mice were untreated. cDNA was synthesized from the livers of each mouse and triplicate PCRs were performed on each sample of cDNA. The mean CT (treated mice) from the triplicate PCRs for the target and internal control gene are 27.2, 28.0, 27.8 (target gene) and 24.2, 24.7 and 24.9 (internal control gene). The mean CT (untreated mice) from the triplicate PCRs for the target and internal control gene are 22.8, 23.0, 22.4 (target gene) and 24.8, 25.0 and 24.7 (internal control gene). What are the mean ± s.d. and the coef?cient of variation (CV) for the relative amount of target gene in both groups of mice? What is the difference in expression of the target gene in the liver of the treated mice compared to the untreated mice?