In miRDeep you can create a config file containing multiple samples.
For instance you can make a config.txt containing the following lines (filename and 3 letter code):
and you could call mapper.pl as follows (example):
mapper.pl config.txt --d --e --h --i --j --l 17 --m --p genome.index --s
reads.fa --t reads vs genome.arf -v --o 4 --q
This is the "official" approach to this and this is definitely the way to go for quantification and for miRNA DE.
You could also make a config.txt only with the WTs and another with the KOs. This depends on your study design actually. I'll paste below the relevant passage from the online help.
PS the relevant text from mirDeep2 doc
The user has sequencing data from different samples e.g. different cell-types. A config.txt file has to be created in which each line designates file locations and a unique 3 letter code. For instance:
The user wishes then to pool these files and use the generated files reads.fa and reads_vs_genome.fa for the miRDeep2 analysis.
mapper.pl config.txt -d -c -i -j -l 18 -m -p genome_index -s reads.fa -t reads_vs_genome.arf
Since the reads_vs_genome.arf still contains the 3 letter code for each read mapped to genome the user can then later on dilute the contribution of the different samples to a predicted or known miRNA. It can also be used for example to define 'high confident' predictions if the results are filtered for miRNAs that have sequencing evidence from at least two samples.
modified 5.8 years ago
5.8 years ago by
IV • 1.2k