I'm interested in generating a fasta file from Ilumina paired reads of my wild type strain. I have an NCBI reference genome I can assemble against and I have already uploaded my 2 reads (using the NCBI reference as my genome), used fastq groomer, aligned them with bowtie and generated my pileup. What I think I want to do though is to extract from the pileup a consensus fasta file (using 20 or something as a quality cutoff threshold) and then used that as my new reference genome to align 2 mutant strains against that are from that genetic background. How can I do that in Galaxy? I don't see any way to go from pileup to fastaq or pileup to fasta using some sort of cutoff. I think I can use samtools pileup2fq based on web posts (and then used something else to make a fasta file), but I would like to do all my analysis in galaxy. Is this possible? Any help appreciated, I am new with Galaxy. -John