fastx_toolkit can't work

[letusgo]

when i use fastx_toolkits ,just like fastq_quality_filter:

$ fastq_quality_filter -q 5 -p 50 -i ./SampleS_1.fq -o ./SampleSfilter_1.fq

unfortunately，it returns as：

fastq_quality_filter: Invalid quality score value (char '#' ord 35 quality value -29) on line 4

how can I solve it ?

[Alex Renwick]

The program is using solexa quality scores (starting at 64) instead of sanger (starting at 33). According to the fastx website, support for sanger quality scores was added in version 0.0.13. Which version are you running?

[dfjenkins3]

Try the command line option -Q 33 to indicate Sanger phred scores. That option has worked for me in the past

[letusgo]

The program is using solexa quality scores (starting at 64) instead of sanger (starting at 33). According to the fastx website, support for sanger quality scores was added in version 0.0.13. Which version are you running?

i use the version 0.0.13,but it doesn't work too,how can i convert sanger fastq to illumina fastq format?

[letusgo]

Try the command line option -Q 33 to indicate Sanger phred scores. That option has worked for me in the past

i add this option ,but another error is happened like:

fastq_quality_filter: Invalid quality score value (char 'J' ord 74 quality value 41) on line 4

[sisch]

Can you post a few (12) lines of your FastQ file?

[mgogol]

I always thought it was -Q33... Does -Q 33 work?

[edit] Oh, yes, it does.

[letusgo]

can you post a few (12) lines of your fastq file?

@hwi-st298:170:81mg3abxx:1:1101:1349:2455 1:n:0:ttaggc
ctggttcagccttcttaggctcttcttctgtttccttacctcagatcggaagagcacacgtctgaactccagtcacttaggcatttcgtatgacgtatgt
+
cccfffffghghfhgiijjfhegi?fhiihfchggggce[email protected]@@e;7=a;aadd>ccbcca################
@hwi-st298:170:81mg3abxx:1:1101:1372:2461 1:n:0:ttaggc
agacggtgttgcataagctgttgtatctgttgctgttgaaactgaaactgctgctgttgctgcttttgttttttagatcggcagggcacacgtgtgcgac
+
@@@[email protected]<gaeehggg@@ehafhiiibgg<?ff<?<bf<[email protected]?be<<3?88bf#######################################
@hwi-st298:170:81mg3abxx:1:1101:1866:2486 1:n:0:ttaggc
gtcactttgtacctcttctcaaattaggaaaatacgaagttcgaggaacgcctttcgttctctttttttcttggtcggcagggtcagggcctttcttgct
+
@@cdffffdddhhegeggeegeh>hhhiigiij>ffcgggeghid0?dfdfggiig1=;;ceggcac<<[email protected]@>>>;??>95(9<(38<?<a#######

[kmcarr]

i add this option ,but another error is happened like:

fastq_quality_filter: Invalid quality score value (char 'J' ord 74 quality value 41) on line 4

letusgo,

Are you sure that you are using v. 0.0.13? See this response to a thread about this issue from last year. (Hint, try searching the forum first, your question may already have been answered.) Using v. 0.0.13 and the '-Q 33' option together should work. If it doesn't then your FASTQ file may be malformed.

[letusgo]

I always thought it was -Q33... Does -Q 33 work?

[edit] Oh, yes, it does.

unfortunately,it doesn't work for my fastq files,then i convert the sanger fastq to illumina fastq.when i run fastq_quality_filter,another error :

fastq_quality_filter: Invalid quality score value (char 'i' ord 105 quality value 41) on line 4

[kmcarr]

@hwi-st298:170:81mg3abxx:1:1101:1349:2455 1:n:0:ttaggc
ctggttcagccttcttaggctcttcttctgtttccttacctcagatcggaagagcacacgtctgaactccagtcacttaggcatttcgtatgacgtatgt
+
cccfffffghghfhgiijjfhegi?fhiihfchggggce[email protected]@@e;7=a;aadd>ccbcca################
@hwi-st298:170:81mg3abxx:1:1101:1372:2461 1:n:0:ttaggc
agacggtgttgcataagctgttgtatctgttgctgttgaaactgaaactgctgctgttgctgcttttgttttttagatcggcagggcacacgtgtgcgac
+
@@@[email protected]<gaeehggg@@ehafhiiibgg<?ff<?<bf<[email protected]?be<<3?88bf#######################################
@hwi-st298:170:81mg3abxx:1:1101:1866:2486 1:n:0:ttaggc
gtcactttgtacctcttctcaaattaggaaaatacgaagttcgaggaacgcctttcgttctctttttttcttggtcggcagggtcagggcctttcttgct
+
@@cdffffdddhhegeggeegeh>hhhiigiij>ffcgggeghid0?dfdfggiig1=;;ceggcac<<[email protected]@>>>;??>95(9<(38<?<a#######

O.K. I missed this before. Clearly your quality lines are malformed; are these after you converted them? You have ASCII values ranging from 35 (#) to 105 (i). No valid FASTQ files for Illumina reads uses a range this wide.

[letusgo]

O.K. I missed this before. Clearly your quality lines are malformed; are these after you converted them? You have ASCII values ranging from 35 (#) to 105 (i). No valid FASTQ files for Illumina reads uses a range this wide.

they come from a seqencing company,i didn't convert them.but when i convert them to Illumina fastq,their ASCII values is also malformed,what can i do?

[arvid]

they come from a seqencing company,i didn't convert them.but when i convert them to Illumina fastq,their ASCII values is also malformed,what can i do?

Ask the company what they did to create such data..?

[ishmael]

Just curious, I googled the unique machine id: hwi-st298.
And I found this:
http://seqanswers.com/forums/showthread.php?t=15928
It looks that the company triggered the malformation not the first time.
Which company it is? BGI?