Awesome! That worked very well, thanks a lot.

P

On Wed, Jun 3, 2015 at 4:26 PM, Heng Li <[email protected]> wrote:

> Bwa-mem -C simply copies FASTQ header comments to output SAM. Therefore,
> the comment must be in the SAM tag format. In your case, you should change
> the FASTQ header to (note the red part):
>
> @HWI-D00595:74:C6DWYANXX:6:1101:1230:2158 cs:Z:
> *1:N:0:TCCGGAGACGCTCTGA:CUSTOM:1:126:14:9.0:14:100:0:252:42:7:7:42:1.59:ATATGTATACATAT:.:.:126:1.0*
>
> ATATGTATACATATATATGTATACATATATATGTATACATATATATGTATACATATATATGTATACATATATATGTATACATATATATGTATACATATATATGTATACATATATATGTATACATAT
> +HWI-D00595:74:C6DWYANXX:6:1101:1230:2158 1:N:0:TCCGGAGACGCTCTGA
>
> /</<<F<FFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFF<FFFFFFFF/FFFFFFFFFFFFF/BFFFFFFFFFFFFBBFBF<BFFF<B/B/FFF<BBFF#
> @HWI-D00595:74:C6DWYANXX:6:1101:1149:2167 cs:Z:
> *1:N:0:TCCGGAGACGCTCTGA:CUSTOM:4:124:16:6.9:18:81:14:123:45:4:4:44:1.47:TATATATCATATATATGA:ATG:AT:126:0.488095238095*
>
> ATGTATATACATATATATGAATATATATTCATATATATGTATATACATATATATGTATATACATATATATGAATATATATATATGTATATACATATATATATATGAATATATATTCATATATATAT
> +HWI-D00595:74:C6DWYANXX:6:1101:1149:2167 1:N:0:TCCGGAGACGCTCTGA
>
> BBBBBFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFBFBFFFFFFFFFFF/FFFFFF</BFFFFFFFFFF//<BFFFFFFF/<<BBFFFFFF
>
>
> Heng
>
> On Jun 3, 2015, at 10:19 AM, Piet Jones <[email protected]> wrote:
>
> Hi,
>
> I am in the process of analysing a set of fastq files (paired-end, human).
> I annotate the reads with information, which is basically a long string of
> numbers and short sequences, eg:
>
> @HWI-D00595:74:C6DWYANXX:6:1101:1230:2158
> *1:N:0:TCCGGAGACGCTCTGA:CUSTOM:1:126:14:9.0:14:100:0:252:42:7:7:42:1.59:ATATGTATACATAT:.:.:126:1.0*
>
> ATATGTATACATATATATGTATACATATATATGTATACATATATATGTATACATATATATGTATACATATATATGTATACATATATATGTATACATATATATGTATACATATATATGTATACATAT
> +HWI-D00595:74:C6DWYANXX:6:1101:1230:2158 1:N:0:TCCGGAGACGCTCTGA
>
> /</<<F<FFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFF<FFFFFFFF/FFFFFFFFFFFFF/BFFFFFFFFFFFFBBFBF<BFFF<B/B/FFF<BBFF#
> @HWI-D00595:74:C6DWYANXX:6:1101:1149:2167
> *1:N:0:TCCGGAGACGCTCTGA:CUSTOM:4:124:16:6.9:18:81:14:123:45:4:4:44:1.47:TATATATCATATATATGA:ATG:AT:126:0.488095238095*
>
> ATGTATATACATATATATGAATATATATTCATATATATGTATATACATATATATGTATATACATATATATGAATATATATATATGTATATACATATATATATATGAATATATATTCATATATATAT
> +HWI-D00595:74:C6DWYANXX:6:1101:1149:2167 1:N:0:TCCGGAGACGCTCTGA
>
> BBBBBFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFBFBFFFFFFFFFFF/FFFFFF</BFFFFFFFFFF//<BFFFFFFF/<<BBFFFFFF
>
> I have highlighted the annotation in bold, part of it is the standard
> annotation such as read number (1 vs 2) and barcode. Now, when I align this
> with bwa mem I use the "-C" flag, this prints this flag to the samfile
> after each alignment, which is great, the problem is that samtools
> complains about this format:
>
> -bash-3.2$ samtools view test_1_annotated.sam
> [E::sam_parse1] unrecognized type
> [W::sam_read1] parse error at line 95
> [main_samview] truncated file.
>
> Removing the annotation makes samtools run without an issue. I know the
> annotation violates the samtools format according to the docs. So my
> question is, is there some other way that I can incorporate this annotation
> and tug it along with my analysis further downstream (such as filtering
> using samtools etc)?
>
> Kind Regards,
> Piet Jones
>
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