Try 'make clean' to replace old objects and dependencies that were generated by the prior makes.
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From what you originally wrote, there's no reason for it to try to go to a non-existent /home/lab/Desktop/JL directory. There's also no reason I've ever seen to edit the Makefile for samtools aside from changing the LIBCURSES value (if needed). I'm guessing that there's something weird with your Makefile. You might try just deleting the samtools directory and redownloading it (from the samtools page on sourceforge and no where else) so you know nothing is screwy. Then, (1) extract the files, (2) cd into the resulting directory and (3) type make without editing any files. You mentioned using Ubuntu earlier and I can confirm that this works correctly under Ubuntu.
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So, I've tried re-downloading both samtools and ubuntu several times. The simple directions to just go into the folder and command "make" is simply not working. And when I get tricky (changing the makefile, downloading my own c++, nsurses, or zlib, trying it as a different user ect.) but it still won't work. I feel as if I've exhausted my google search results. I even contacted the samtools people, who told me to download and try again. This I have done many many times. What could I be missing?
Thank you for your continuing comments. I can't tell you how much I appreciate your efforts on my behalf.
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Here's what I did to download and compile a new version of samtools:
The wget command goes through a few redirects before downloading. The make process takes about 10 seconds. After that I can use the samtools mpileup command:Code:wget -O samtools-0.1.18.tar.bz2 http://sourceforge.net/projects/samtools/files/samtools/0.1.18/samtools-0.1.18.tar.bz2/download tar xjf samtools-0.1.18.tar.bz2 cd samtools-0.1.18 make make razip
At what point does this process fail for you?Code:[server samtools-0.1.18]$ ./samtools mpileup Usage: samtools mpileup [options] in1.bam [in2.bam [...]] Input options: -6 assume the quality is in the Illumina-1.3+ encoding -A count anomalous read pairs -B disable BAQ computation -b FILE list of input BAM files [null] -C INT parameter for adjusting mapQ; 0 to disable [0] -d INT max per-BAM depth to avoid excessive memory usage [250] -E extended BAQ for higher sensitivity but lower specificity -f FILE faidx indexed reference sequence file [null] -G FILE exclude read groups listed in FILE [null] -l FILE list of positions (chr pos) or regions (BED) [null] [...etc...]
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THANK YOU everyone for your help!! I did get this to work. In case some poor student is struggling with the same thing I was, here's what finally worked:
1) I deleted the samtools folders I had.
2) used command:
sudo apt-get install samtools
3) Now, I need to figure out where it went, so I did
dpkg -s samtools
It showed-- samtools: /usr/bin/samtools /usr/lib/samtools /usr/share/samtools /usr/share/man/man1/samtools.1.gz
4) In usr/bin, the executable is locked. To unlock I did
cd ../
cd usr
cd bin
sudo chmod a+rwx samtools
5) manually move the files I'm working with (the reference.fasta and the output.sorted.bam) to the usr>bin folder
6) try command in usr>bin folder
sudo samtools mpileup -f reference.fasta output.sorted.bam >output.mpileup
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by GATTACATLove this - good data definitely starts from good input, and poor input can only give relatively poor data. I particularly like the mention of Nanodrop/absorbance based methods for quantification. It's such a toss up if you'll get an accurate reading or what amounts to a randomly generated number, and a lot of library/sequencing related issues can be traced back to poor quant.
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Channel: Articles
07-01-2026, 11:43 AM -
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by SEQadmin2
I’m not a sequencing expert. I’m a purification scientist who uses NGS to evaluate workflows my group develops. With this perspective, we think about the sample first and the NGS workflow second. The sequencer is an exceptionally honest reporter, but it can only report on what you give it, so whether you get clean, interpretable data from an NGS workflow is largely determined before you begin.
Here are nine questions we think about, in roughly the order they matter, before...-
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