Ahhh, new samtools sort requires either the -o or -O flag.  See the man page.

On Mon, 27 Jul 2015, Thomas W. Blackwell wrote:

>
> Must always sort before indexing.  Use 'samtools sort sample.bam > 
> sorted.bam'.
>
> On Mon, 27 Jul 2015, Komal Rathi wrote:
>
>> Hi everyone,
>> 
>> I am using samtools 1.2/htslib 1.2.1.
>> 
>> The following is my bam file - mapped to mm10:
>> samtools view -h sample.bam | less
>> 
>> @HD     VN:1.0  SO:unsorted
>> @PG     ID:GSNAP        PN:gsnap        VN:2014-12-31   CL:gsnap -d 
>> mm10_gmap --batch=5 --nthreads=10 --format=sam 
>> reads_bothclipped_unaligned.fa
>> @SQ     SN:chr1 LN:195471971
>> @SQ     SN:chr2 LN:182113224
>> @SQ     SN:chr3 LN:160039680
>> @SQ     SN:chr4 LN:156508116
>> @SQ     SN:chr5 LN:151834684
>> @SQ     SN:chr6 LN:149736546
>> @SQ     SN:chr7 LN:145441459
>> @SQ     SN:chr8 LN:129401213
>> @SQ     SN:chr9 LN:124595110
>> @SQ     SN:chr10        LN:130694993
>> @SQ     SN:chr11        LN:122082543
>> @SQ     SN:chr12        LN:120129022
>> @SQ     SN:chr13        LN:120421639
>> @SQ     SN:chr14        LN:124902244
>> @SQ     SN:chr15        LN:104043685
>> @SQ     SN:chr16        LN:98207768
>> @SQ     SN:chr17        LN:94987271
>> @SQ     SN:chr18        LN:90702639
>> @SQ     SN:chr19        LN:61431566
>> @SQ     SN:chrX LN:171031299
>> @SQ     SN:chrY LN:91744698
>> @SQ     SN:chrM LN:16299
>> HWI-D00712:1134:C6NMYANXX:5:1101:11882:3444     4       *       0       0 
>> *       *       0       0       ACACGCAGCAACCCATCGTT    *       XO:Z:NM
>> HWI-D00712:1134:C6NMYANXX:5:1101:10181:1990     4       *       0       0 
>> *       *       0       0       GTTGGGGTCGTAGCCTCGT     *       XO:Z:NM
>> HWI-D00712:1134:C6NMYANXX:5:1101:5509:2135      4       *       0       0 
>> *       *       0       0       GCCCACCTGTTAGTGTTGT     *       XO:Z:NM
>> HWI-D00712:1134:C6NMYANXX:5:1101:4268:5419      4       *       0       0 
>> *       *       0       0       GTCGATCCGGGCTTTCGTT     *       XO:Z:NM
>> HWI-D00712:1134:C6NMYANXX:5:1101:10454:4937     4       *       0       0 
>> *       *       0       0       TCTGCGAACCAACACG        *       XO:Z:NM
>> HWI-D00712:1134:C6NMYANXX:5:1101:4687:5341      4       *       0       0 
>> *       *       0       0       TACCTCCTAGTCTTGGTTT     *       XO:Z:NM
>> HWI-D00712:1134:C6NMYANXX:5:1101:18702:6367     4       *       0       0 
>> *       *       0       0       TTACCTTCACATGATCTGCT    *       XO:Z:NM
>> HWI-D00712:1134:C6NMYANXX:5:1101:4506:7580      4       *       0       0 
>> *       *       0       0       CAGTCACAATTACCGGT       *       XO:Z:NM
>> HWI-D00712:1134:C6NMYANXX:5:1101:16573:3163     4       *       0       0 
>> *       *       0       0       CTTTATCAATAGAGAGT       *       XO:Z:NM
>> HWI-D00712:1134:C6NMYANXX:5:1101:15871:7576     4       *       0       0 
>> *       *       0       0       ACTTCATGCTCATTGGTTT     *       XO:Z:NM
>> HWI-D00712:1134:C6NMYANXX:5:1101:16993:8112     4       *       0       0 
>> *       *       0       0       AGGGATGAGCTCACCGT       *       XO:Z:NM
>> HWI-D00712:1134:C6NMYANXX:5:1101:17679:3919     4       *       0       0 
>> *       *       0       0       TAGATCGCTTTGATCCGT      *       XO:Z:NM
>> HWI-D00712:1134:C6NMYANXX:5:1101:20428:8085     4       *       0       0 
>> *       *       0       0       CGAGAGAGATTACCTCCGTT    *       XO:Z:NM
>> HWI-D00712:1134:C6NMYANXX:5:1101:13289:6144     4       *       0       0 
>> *       *       0       0       ACACCGCTCTAAGAGCGT      *       XO:Z:NM
>> 
>> I am trying to index it using:
>> samtools index sample.bam
>> 
>> but it is throwing an error:
>> [E::hts_idx_push] NO_COOR reads not in a single block at the end 1 -1
>> 
>> Any help would be much appreciated!
>> 
>> --Komal
>> 
>>
>> 
>> ------------------------------------------------------------------------------
>> _______________________________________________
>> Samtools-help mailing list
>> [email protected]
>> https://lists.sourceforge.net/lists/listinfo/samtools-help
>> 
>

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