From 49b9bc665af1b9c52ec133952cdddf9632c898e3 Mon Sep 17 00:00:00 2001 From: AshKernow Date: Thu, 3 Oct 2024 10:03:41 +0100 Subject: [PATCH] Minor formatting changes --- Markdowns/08_DE_analysis_with_DESeq2.Rmd | 16 +++++++++------- Markdowns/08_DE_analysis_with_DESeq2.html | 16 +++++++++++++--- Markdowns/08_DE_analysis_with_DESeq2.pdf | Bin 6003344 -> 6003723 bytes 3 files changed, 22 insertions(+), 10 deletions(-) diff --git a/Markdowns/08_DE_analysis_with_DESeq2.Rmd b/Markdowns/08_DE_analysis_with_DESeq2.Rmd index 2280c97..3e1a770 100644 --- a/Markdowns/08_DE_analysis_with_DESeq2.Rmd +++ b/Markdowns/08_DE_analysis_with_DESeq2.Rmd @@ -113,8 +113,9 @@ and day 11. ## Exercise 1 > This time create and investigate the model matrix for the variable "Status". -> 1. Create a model formula to investigate the effect of “Status” on gene expression. -> 2. Look at the model matrix and identify which is the reference group in your model. +> +> 1. Create a model formula to investigate the effect of “Status” on gene expression. +> 2. Look at the model matrix and identify which is the reference group in your model. The intercept has been set automatically to the group in the factor that is alphabetically first: `Infected`. @@ -351,11 +352,12 @@ ddsObj.filt <- ddsObj.raw[keep, ] > called `results.additive` > > Questions: -> a) How many coefficients are there in the additive model? -> b) What is the reference group in the additive model? -> c) What contrasts could we perform using this model? -> d) What contrast does the `results.additive` object represent? -> e) How many genes have an adjusted p-value of less than 0.05 +> +> a) How many coefficients are there in the additive model? +> b) What is the reference group in the additive model? +> c) What contrasts could we perform using this model? +> d) What contrast does the `results.additive` object represent? +> e) How many genes have an adjusted p-value of less than 0.05 ```{r echo = FALSE, message = FALSE, include = FALSE} additive.model <- as.formula(~ TimePoint + Status) diff --git a/Markdowns/08_DE_analysis_with_DESeq2.html b/Markdowns/08_DE_analysis_with_DESeq2.html index 8442c0c..de8f9b6 100644 --- a/Markdowns/08_DE_analysis_with_DESeq2.html +++ b/Markdowns/08_DE_analysis_with_DESeq2.html @@ -269,7 +269,7 @@

Introduction to Bulk RNAseq data analysis

Differential Expression of RNA-seq data

-

Last modified: 30 Sep 2024

+

Last modified: 03 Oct 2024

@@ -337,7 +337,12 @@

The simple model

Exercise 1

-

This time create and investigate the model matrix for the variable “Status”. 1. Create a model formula to investigate the effect of “Status” on gene expression. 2. Look at the model matrix and identify which is the reference group in your model.

+

This time create and investigate the model matrix for the variable “Status”.

+
    +
  1. Create a model formula to investigate the effect of “Status” on gene expression.
    +
  2. +
  3. Look at the model matrix and identify which is the reference group in your model.
  4. +

The intercept has been set automatically to the group in the factor that is alphabetically first: Infected.

It would be nice if Uninfected were the base line/intercept. To get R to use Uninfected as the intercept we need to use a factor. Let’s set factor levels on Status to use Uninfected as the intercept.

@@ -525,7 +530,12 @@

Exercise 3

  • Run the size factor estimation, dispersion estimation and modelling steps using the DESeq command as above.

  • Extract the default contrast using the results command into a new object called results.additive

  • -

    Questions: a) How many coefficients are there in the additive model? b) What is the reference group in the additive model? c) What contrasts could we perform using this model? d) What contrast does the results.additive object represent? e) How many genes have an adjusted p-value of less than 0.05

    +

    Questions:

    +
    a) How many coefficients are there in the additive model?   
    +b) What is the reference group in the additive model?    
    +c) What contrasts could we perform using this model?    
    +d) What contrast does the `results.additive` object represent?   
    +e) How many genes have an adjusted p-value of less than 0.05  

    The default contrast of results function

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