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Alex • 0
@da74b9f0
Last seen 9 days ago
United States

Hi,

My design looks like this ==I have 3 factors:

I'm interested to look at the effects of treatment(infection) on the gene expression in each organ over time. I'm having troubles understanding how to define the reduced model to run DESeq.

here is my code:

dds <- nbinomLRT(dds, full = ~Treatment + Organ + Time + Treatment:Time + Organ:Time, reduced = ~Treatment + Organ + Treatment:Time)

#rlog transformation
rld <- rlog(dds, blind=FALSE)
rld_mat <- assay(rld)


My questions:

1. Are my full and reduced models correct for the intended purpose?
2. Is my code for rlog transformation correct with "blind=FALSE" or should I use "blind=TRUE"?

Thanks!

DESeq2 • 218 views
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@mikelove
Last seen 2 days ago
United States

For questions about how to formulate particular contrasts in R, I recommend working with a local statistician or someone familiar with linear models in R. I have limited time on the support site, and have to reserve my time for software related questions.

It is fine to use blind=FALSE.

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Thanks.

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Hi I was able to get an insight on the design. What is the difference in hypothesis testing between these two designs?

1. nbinomLRT(dds2, full = ~Organ + Time + Organ:Time, nbinomLRT(dds, full = ~Organ + Time + Organ:Time, reduced = ~Organ + Time)
2. nbinomLRT(dds2, full = ~Organ + Time + Organ:Time, reduced = ~1)


when I run both, there is a difference in the number of genes that are differentially expressed.

summary(res_LRT) ====1st design!

out of 1359 with nonzero total read count
LFC > 0 (up)       : 11, 0.81%
LFC < 0 (down)     : 8, 0.59%
outliers [1]       : 0, 0%
low counts [2]     : 869, 64%
(mean count < 2)
[1] see 'cooksCutoff' argument of ?results
[2] see 'independentFiltering' argument of ?results


and

summary(res_LRT2) ====2nd design!

out of 1359 with nonzero total read count
LFC > 0 (up)       : 56, 4.1%
LFC < 0 (down)     : 67, 4.9%
outliers [1]       : 0, 0%
low counts [2]     : 527, 39%
(mean count < 1)
[1] see 'cooksCutoff' argument of ?results
[2] see 'independentFiltering' argument of ?results

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For questions about statistical analysis and interpretation, I recommend working with a local statistician or someone familiar with linear models in R.