CASAVA 1.8.2 configureBltoFastq error

[redindian]

Hello to everyone from a new user -I am trying to learn Casava 1.8.2. I am working with a single read 50 bp read and 7bp index run(6 base index--some samples are TruSeq RNA and some lanes have small RNA indices). After running the configureBcltoFastq script I ran make (assuming it does read 1 and read2 for index?). It sttarts but then stops at one of teh lanes giving an error that it "Failed to parse the options:
*** sample-dir not valid: number of directories must match the number of barcodes ***" I did notice that it is reporting all 6 multiplex barcodes and an additional one which it calls undetermined.

Any input? Thanks much!

[sklages]

Originally Posted by redindian

Hello to everyone from a new user -I am trying to learn Casava 1.8.2. I am working with a single read 50 bp read and 7bp index run(6 base index--some samples are TruSeq RNA and some lanes have small RNA indices). After running the configureBcltoFastq script I ran make (assuming it does read 1 and read2 for index?). It sttarts but then stops at one of teh lanes giving an error that it "Failed to parse the options:
*** sample-dir not valid: number of directories must match the number of barcodes ***" I did notice that it is reporting all 6 multiplex barcodes and an additional one which it calls undetermined.

Any input? Thanks much!

How does your sample sheet look like? How did you start configureBcltoFastq?
We need a bit more information ...

cheers,
Sven

[dlawrence]

*** sample-dir not valid: number of directories must match the number of barcodes ***"

I just had the same problem.

Short answer: don't put spaces in the SampleSheet.csv fields.

Long answer:

Casava runs in multiple threads causing any errors to be interleaved with debug messages in the log... to work out what lane the problem was, I looked in the Temp directory, and found the one without a XXX_demux_summary.xml file.

In the SampleSheet.csv for that lane I had 5 samples: "Mut F5 Rep1","Mut F5 Rep3" etc. Looking in the lane's directory in Temp/ there was just 1: "Sample_Mut".

So, we've found the source of the error (num directories not equaling num sample sheet samples)

My guess is Casava is not quoting spaces properly in one of the scripts, and this caused the lane samples to overwrite each other, then the directory check failed.

I edited the SampleSheet, changed spaces to dashes, reran and everything was fine. I'll email Illumina and lodge a bug report to fix their quoting.

[sklages]

That is written in "CASAVA v1.8.2 User Guide", p. 31.

We disallowed whitespace and some other characters in sequencing sample names since our first ABI 373 sequencers .. and we still stick to that ;-)

[dlawrence]

Yeah, but it would be good to die immediately with a nice message, not cryptically 15 CPU hours later

[sklages]

Originally Posted by dlawrence

Yeah, but it would be good to die immediately with a nice message, not cryptically 15 CPU hours later

I totally agree, .. but that's the typical UI from Illumina. All of their (unix) software behaves this way; that's partly due to using the 'make' system for processing the data. OLB is funny for error tracking as well ..