I have methylation data from ONT sequencing expressed as percentage of methylation and/or number of reads. I don't know if I'm supposed to normalize the data and how to handle replicates. Some ideas: I have data in triplicate. I want to normalize the data (quantile normalization is the preferred option) and/or collapse data to have 1 file/dataset from 3 replicates (doing weigthed average maybe). So 1 idea would be just to perform weighted average of each methylation value doing [%M*NR] where %M is the percentage of methylated reads and NR is number of reads, per each methylated position. In this case I will lose the meaning of the percentage value in the downstream analyses because I will have a wheigted value. otherwhise I can perform quantile normalization on the number of reads (total and methylated) and then calculate percentage of methylated reads. I would like the opinion of somewone with a better statistical background than me ;) thansk for your kind help!
%M-S1 NR-S1 %M-S2 NR-S2 %M-S3 NR-S3
Meth1 20 60 15 54 41 12
Meth2 40 14 78 52 13 65
Meth3 12 94 73 19 37 70
Meth4 36 77 69 14 26 74