Question: Issue With CRAM -> BAM -> FASTQ Conversion
0
gravatar for denis.k
12 months ago by
denis.k10
denis.k10 wrote:

Please help! I am trying to obtain fastq files from the GDSC, all we have in the lab is CRAM files. Unfortunately, the reference genome seems to not exist when pulled from an online source. I have attempted to use the Samtools code which allows me to put in my own reference but I get an error message

code used:

 samtools view -b  -T /GRCh37.p13.genome.fa -o /EGAR00001252191_13305_1.bam /EGAR00001252191_13305_1.cram

error message:

[W::find_file_url] Failed to open reference "https://www.ebi.ac.uk/ena/cram/md5/1b22b98cdeb4a9304cb5d48026a85128": Protocol not supported
[E::cram_get_ref] Failed to populate reference for id 0
[E::cram_decode_slice] Unable to fetch reference #0 10507..19965
[E::cram_next_slice] Failure to decode slice
[main_samview] truncated file.

Samtools v1.9, OS: Ubuntu 18.04.2 LTS

Any ideas? Thank you in advance!

view cram bam fastq samtools • 992 views
ADD COMMENTlink modified 11 months ago by jkbonfield390 • written 12 months ago by denis.k10

I assume this data is coming from European Genome Phenome Archive? You should email datasharing at sanger.ac.uk to get assistance with reference.

ADD REPLYlink modified 12 months ago • written 12 months ago by genomax85k

Hello,

how have you installed samtools? Is htslib installed as well?

Related: https://github.com/samtools/samtools/issues/857

fin swimmer

ADD REPLYlink written 12 months ago by finswimmer13k
3
gravatar for jkbonfield
11 months ago by
jkbonfield390
jkbonfield390 wrote:

Has your copy of samtools (htslib actually) been compiled with curl support? If you just did "make" the answer will probably be no. You need ./configure;make. It's dumb and an annoyance of mine that we have two different build routes that by default choose two different combinations of features. Argh. This is almost certainly what the "protocol not supported" means as without curl it won't be able to handle https; only http.

If you do indeed have a curl capable samtools, then is it maybe failing due to a firewall rule? You can use htslib's "hts_file -vvvvv -c in.cram > /dev/null" to read a CRAM file with maximum verbosity enabled, which may shed some light on the manner. (Sadly samtools itself doesn't have any way to enter htslib debug mode, unless you attach a debugger to it.)

ADD COMMENTlink written 11 months ago by jkbonfield390
1

Thank you! It was a lib curl problem! Turns out that I just needed the --enable lib curl option. Such a silly problem. Thank you for all the help

ADD REPLYlink written 10 months ago by denis.k10
Please log in to add an answer.

Help
Access

Use of this site constitutes acceptance of our User Agreement and Privacy Policy.
Powered by Biostar version 2.3.0
Traffic: 739 users visited in the last hour