RNA-SEQ data processing

Hello All,
I am looking into co-expression analysis of genes at different developmental stages. I am stuck up with few things. I would be grateful if you could help me out.

Basically I want to find genes coexpressed to a set of bait genes in different stages and genes co-expressed in particular stage too. So I thought I should do DEG analysis between stages and co-expression analysis for all stages and then I can get uniquely coexpressed genes in one stage. So I need to do DEG and coexpression. I am using publicly available RNA-SEQ data.

In one dataset I have genes, FPKM values, their annotations and the trinity assembly(fasta). But, I found that using FPKM values for DEG or co-expression analysis is not recommended and I neither can give it as input to existing softwares. So I do not figure out what can I do here.

In the second dataset, I have logCPM values obtained from (limma,voom:Reference-manuscript methods) of genes across different stages. They range from -5 to 12. How do I find DEG with this? Can I directly use them to take cutoff and what cutoff? Cause I do not have any other details.

Thanks in advance!

rna-seq

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updated 7.8 years ago by ng8423002 •

• written 7.8 years ago by meenakshikapase •

2 answers

You are correct. FPKM is no more considered suitable as mentioned in the post here

FPKM not suitable for DE?

What I will suggest is to use tools which use raw read counts. check out this post

differentially expression analysis using raw read counts

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written 7.8 years ago by lakhujanivijay

RNA-sequencing (RNA-seq) has a wide variety of applications, but no single analysis pipeline can be used in all cases. We review all of the major steps in RNA-seq data analysis, including experimental design, If you are facing any issue regarding with virus and malware, you can take help from kaspersky support number, http://kasperskysupportnumber.co.uk/. thanku good luck.

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written 7.8 years ago by ng8423002 •