Question: Differential Expression Analysis with monocle and batch effect correction
1
gravatar for Poorya Parvizi
4 weeks ago by
Middle East Technical University
Poorya Parvizi30 wrote:

Hi,

I would like to run differential expression analysis on my single-cell data. My data contains of 5 groups which sequenced in 3 batches. Each batch contains all of the groups. Before the differential expression analysis with monocle (I should run it with monocle) I have tried to eliminate batch effect from the data. I used Seurat's ScaleData function (vars.to.regress argument) to regress out the batch effect. However, the output of this function is in scale format which contains negative values in it.

Can I use scale values in differential expression analysis, specifically monocle?

I also tried limma's removeBatchEffect function, which also gives negative values. What is the best way to regress out the batch effect before the monocle differential expression?

ADD COMMENTlink modified 4 weeks ago by Kevin Blighe21k • written 4 weeks ago by Poorya Parvizi30
1
gravatar for Kevin Blighe
4 weeks ago by
Kevin Blighe21k
University College London Cancer Institute
Kevin Blighe21k wrote:

I do not believe negative values will help differential expression analysis in any way. Monocle actually provides functionality for dealing with things like batch. In many of the functions, there is a parameter called residualModelFormulaStr, which allow you to list any covariates for which the statistical modelling should be adjusted.

residualModelFormulaStr

A model formula string specify effects you want to exclude when testing for cell type dependent expression

So, for example, for a differential expression analysis, use:

differentialGeneTest(cds, fullModelFormulaStr = " ~ condition",
reducedModelFormulaStr = " ~ Batch", relative_expr=TRUE, cores=4)

Take a look at the Manual and Tutorial.

Kevin

ADD COMMENTlink modified 7 days ago • written 4 weeks ago by Kevin Blighe21k

Thank you Kevin. I will try it.

ADD REPLYlink written 4 weeks ago by Poorya Parvizi30
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