04 April 2018 4 3K Report

Hello everyone.

Right now i am working on differential expression of miRNA using next generation sequencing. analysis post sequencing was done using "Small rna analysis" pipeline in a integrated analysis software. I have disagreement with my supervisor regarding the results of dysregulated miRNA in terms of normalization method used. I use small rna analysis pipeline, because it was explained in the software about this pipeline if i want to analyse miRNA differential expression. Many journals also use this pipeline to find differential expression of miRNA. Regarding normalization method, i also read that method of normalization for RNA seq can be used for small RNA seq, for example DESeq/edgeR, TMM etc. but my supervisor need more assurance because he wanted normalization method specifically for miRNA sequencing. Since i am new in this field, i need some confirmation from experienced people in this forum.

1. Is it right for me to think that miRNA sequencing is the same as small RNA sequencing, and almost the same with RNA sequencing (just different on enriched small RNA annotation)? if

2. if previous question's answer is yes, so is it okay to use normalization method usually done for RNA seq,for example DESeq etc, and what is the reason for this?

if the answer is no, so what is actually the difference between those sequencing, and what kind of normalization can be used specifically for mirNA sequencing?

Looking forward for any link or elaboration from experienced people here in this forum.

Regards,

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