I am working with RNASeq data and completely new in it. I have already performed differential expression analysis using DESeq2 and identified differentially expressed genes.
Now with the identified differentially expressed genes, I want to perform clustering to identify which genes works in groups for their biological activities.
Please suggest me
1) Which normalization technique is the most suitable right now for RNASeq data along with links/code for performing that normalisation technique.
2) Is log2 transformation is to be accompanied with the normalisation as it was the case in Microarray analysis?
3) In DESeq2 tutorials, there is vst and rlog transformation, is it necessary to perform any one of those along with other normalization / preprocessing steps?
3) Please provide me link for tutorials or code for performing the most suitable normalisation technique and any other necessary steps (if any) before I can apply any clustering techniques like hierarchical clustering with those preprocessed gene expressions of the differentially expressed genes identified using DESeq2.
4) Whether there is any difference in Differential Expression analysis techniques and Normalization procedure for single cell RNASeq (scRNASeq) data and RNASeq data?
5) How do I ensure whether a NCBI geo series data is just RNASeq or scRNASeq? I mean is there any distinct text written in NCBI all accession series home page that it's RNASeq or scRNASeq?
Thanks in advance.