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Dual-RNA seq with bacteria and fungi- can you split co-culture libraries for DGE comparisons?

Hi all,

I am new to RNA seq analysis and am looking for some expertise.

I am trying to plan how to analyse dual-RNA seq data that I have been given. The data is not normal human v pathogen data but is combined fungi and bacterial samples.

We are interested in genes which are DGE in fungi in response to inhibitory bacteria and non-inhibitory bacteria. As well as DGE in inhibitory and non- inhibitory bacteria in response to being co-cultured with fungi.

The Illumina pe libraries I have (all in triplicate) are:

-Fungi only -inhibitory bacterium only -non inhibitory bacterium only -co-cultured fungi and inhibitory bacterium -co-cultured fungi and non-inhibitory bacterium only

Is it possible to split libraries from each co-culture samples into separate species and compare DEGs with the bacteria and fungi only control samples? So:

-Fungi only vs fungi from co culture with inhibitory bacteria -inhibitory bacterium only v inhibitory bacterium only from co culture with inhibitory bacteria

And

-Fungi only vs fungi from co culture with non-inhibitory bacteria -non-inhibitory bacterium only v non-inhibitory bacterium only from co culture with inhibitory bacteria

Or is it only possible to do dual RNAseq analysis with co-cultured fungi and inhibitory bacterium and co-cultured fungi and non-inhibitory bacterium only samples?

I’d be grateful for any advice you could give me!

illumina r rna-seq dge transcriptomics

I am trying to plan how to analyse dual-RNA seq data that I have been given. The data is not normal human v pathogen data but is combined fungi and bacterial samples.

How were these libraries prepared? Different prep methods are needed for pro- and eukaryotic libraries.

Is it possible to split libraries from each co-culture samples into separate species and compare DEGs with the bacteria and fungi only control samples?

For the question above answer is yes as long as you have good reference genomes available you should be able to bin/split the reads using bbsplit.sh from BBMap suite.

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