Dear galaxy help, Devon new user making good headway on galaxy cloud here at Case Western Reserve university in Cleveland Ohio. I have a small project. 10 human samples. I did a custom capture of exons from 171 genes. Total target size is ~ 0.5 Gb. PE reads from Illumina HighScan. See attached screenshots. I uploaded one fastq file (1.2Gb). I succeeded in doing fastQC, groomer, trimmer, allignment with Bowtie, SAM to BAM, mpileup. Now, I want to convert to .vcf. for downstream work. I got stuck at bcf tools ( see second slide). It is as if bcf tools doesn't recognize the output from mpileup. Note how the option windows in bcf tools are all collapsed. I used default parameters throughout the workflow. Any Ideas. Patrick -- Patrick Leahy, Ph.D. *Asst. Professor, General Medical Sciences- Oncology* Scientific Coordinator, Integrated Genomics Shared Resource (IGSR) Scientific Managing Director Microarray Core Facility Director, Laser Capture Microdissection Service Case Western Reserve University Room 3541 Wolstein Research Building 2103 Cornell Rd Cleveland, OH 44106 t: 216 368 0761 f: 216 368 8919 http://cancer.case.edu/members/gms_onc/leahy.html