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11. Transcriptomics II

PP
by Pia P.

Which statements regarding the comparisons of expression values are correct?

1) The main reason why expression values are log-transformed is to meet the assumption of a t-test

2) If one compares the mean of a sample of 10 measurements to a reference value one can use a t-distribution with 9 degree of freedoms to determine a significant deviation

3) A paired t-test is also called non-parametric test

4) Non-overlapping error bars representing the standard error of the mean always imply a significant (p<5%) difference between the means of the two populations

5) Non-overlapping error bars representing the 95% C.I. of the mean always imply a significant (p<5%) difference between the means of the two populations

6) An ANOVA that compares two groups for one factor gives exactly the same p-value as a two-sample t-test

Which statements regarding the comparisons of expression values are correct?

1) The main reason why expression values are log-transformed is to meet the assumption of a t-test -> wrong; because we analyze relative expression levels; we are interested in fold-changes, not absolute changes

2) If one compares the mean of a sample of 10 measurements to a reference value one can use a t-distribution with 9 degree of freedoms to determine a significant deviation -> correct

3) A paired t-test is also called non-parametric test -> wrong, t-test is parametric because it makes the assumption of a (normal) distribution; non-parametric tests do not make an assumption

4) Non-overlapping error bars representing the standard error of the mean always imply a significant (p<5%) difference between the means of the two populations -> wrong

5) Non-overlapping error bars representing the 95% C.I. of the mean always imply a significant (p<5%) difference between the means of the two populations -> correct

6) An ANOVA that compares two groups for one factor gives exactly the same p-value as a two-sample t-test -> correct

 You have 12 brain samples from Alzheimer patients and 12 age- and sex-macthed controls. You isolate RNA twice from each sample (i.e. make technical replicates) and sequence 48 RNA-Seq libraries in total. You quantify gene expressionlevels for all detectable genes and want to identify genes differently expressed in Alzheimer patients using a two-sample t-test. Which statements are correct?

1) The two technical replicates are not independent since they come from the same individual

2) One could average the expression levels of the two technical replicates per individual and then calculate a t-test with N=12 per group in order to avoid pseudoreplication

3) The technical replicates allow to estimate how much variations is caused by RNA isolation, library preparation and sequencing

4) Since the RNAs were independently isolated one can calculate the t-test assuming N=24 for patients and N=24 per control

5) The variance between technical replicates is expected to be smaller than the variance between two Alzheimer patients (i.e. between biological replicates)

 You have 12 brain samples from Alzheimer patients and 12 age- and sex-macthed controls. You isolate RNA twice from each sample (i.e. make technical replicates) and sequence 48 RNA-Seq libraries in total. You quantify gene expressionlevels for all detectable genes and want to identify genes differently expressed in Alzheimer patients using a two-sample t-test. Which statements are correct?

1) The two technical replicates are not independent since they come from the same individual -> correct

2) One could average the expression levels of the two technical replicates per individual and then calculate a t-test with N=12 per group in order to avoid pseudoreplication -> correct

3) The technical replicates allow to estimate how much variations is caused by RNA isolation, library preparation and sequencing -> correct

4) Since the RNAs were independently isolated one can calculate the t-test assuming N=24 for patients and N=24 per control -> wrong

5) The variance between technical replicates is expected to be smaller than the variance between two Alzheimer patients (i.e. between biological replicates) -> correct

Author

Pia P.

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