Hi guys,

I'm running velocyto on bam files via ssh. One sample was succeeded, but when I started running the other two samples, I saw this error showed up over and over again, at the beginning, I thought something was wrong with my bam files, so I re-transferred them to the server, and changed the memory in the bash file, as these two samples are bigger than the first sample, but I still see the same error:

File "/wynton/home/anderson/yiwang/.local/bin/velocyto", line 11, in <module>
    load_entry_point('velocyto==0.17.17', 'console_scripts', 'velocyto')()
  File "/wynton/home/anderson/yiwang/.local/lib/python3.6/site-packages/click/core.py", line 829, in __call__
    return self.main(*args, **kwargs)
  File "/wynton/home/anderson/yiwang/.local/lib/python3.6/site-packages/click/core.py", line 782, in main
    rv = self.invoke(ctx)
  File "/wynton/home/anderson/yiwang/.local/lib/python3.6/site-packages/click/core.py", line 1259, in invoke
    return _process_result(sub_ctx.command.invoke(sub_ctx))
  File "/wynton/home/anderson/yiwang/.local/lib/python3.6/site-packages/click/core.py", line 1066, in invoke
    return ctx.invoke(self.callback, **ctx.params)
  File "/wynton/home/anderson/yiwang/.local/lib/python3.6/site-packages/click/core.py", line 610, in invoke
    return callback(*args, **kwargs)
  File "/wynton/home/anderson/yiwang/.local/lib/python3.6/site-packages/velocyto/commands/run.py", line 116, in run
    samtools_memory=samtools_memory, dump=dump, loom_numeric_dtype=dtype, verbose=verbose, additional_ca=additional_ca)
  File "/wynton/home/anderson/yiwang/.local/lib/python3.6/site-packages/velocyto/commands/_run.py", line 229, in _run
    results = exincounter.count(bamfile_cellsorted, multimap=multimap)  # NOTE: we would avoid some millions of if statements evaluations if we write two function count and count_with output
  File "/wynton/home/anderson/yiwang/.local/lib/python3.6/site-packages/velocyto/counter.py", line 756, in count
    for r in self.iter_alignments(bamfile, unique=not multimap):
  File "/wynton/home/anderson/yiwang/.local/lib/python3.6/site-packages/velocyto/counter.py", line 251, in iter_alignments
    fin = pysam.AlignmentFile(bamfile)  # type: pysam.AlignmentFile
  File "pysam/libcalignmentfile.pyx", line 742, in pysam.libcalignmentfile.AlignmentFile.__cinit__
  File "pysam/libcalignmentfile.pyx", line 952, in pysam.libcalignmentfile.AlignmentFile._open
  File "pysam/libchtslib.pyx", line 365, in pysam.libchtslib.HTSFile.check_truncation
OSError: no BGZF EOF marker; file may be truncated

I googled, I found ppl had the same error as me, they suggested checking the bam files with samtools, I did, but I can't tell if it's correct or not:

samtools view GW_possorted_genome_bam.bam | tail
A00269:304:H2MKKDSXY:4:2141:9218:28166  4   *   0   0   *   *   0   0   CTGTGCACCCCGAATCTGCCCCTATCCCCTTCTCTGCTTCCTGCACTGCACACCTGCTAGTCCCAGACCCCTCCCCCAAATAAACGTGAGGGTTAAAAGTTAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA  FFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFF:FFFFFFFFFFFFFFFFFF:FFF:FFF:F:F:::F:FFF:FFFF,F:FFFF:FFFFFFFFFFFFFFFFFFF:FFFFFF:FF:F:FFFFFFF:      NH:i:0  HI:i:0  AS:i:91 nM:i:0  uT:A:1  xf:i:0  li:i:0  BC:Z:TTGCCGGA   QT:Z:FFFFFFF:   CR:Z:TTTGTTGTCTTCACGC   CY:Z:FFFFFFFFFFFFFFFF   CB:Z:TTTGTTGTCTTCACGC-1 UR:Z:AGGTTTGGATTG   UY:Z:FFFFFFFFFFFF   UB:Z:AGGTTTGGATTG   RG:Z:test:0:1:H2MKKDSXY:4
A00269:304:H2MKKDSXY:4:2344:12192:34601 4   *   0   0   *   *   0   0   CCCGCCCCTGTCCCCTAGCCGCCGCTCCGGGCTCTGCTCGCCCTCCTACGTTGCGGTCACACCCTTCTCCCTTCGGGGAGACAACGACGGCGGTGGCGGGAGCTTCTCCACGGCCGACCAGCTGGAGATGGTGACCGAGCTGCTGGGAGG  FFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFF::FFFF:FFFFFFFFF:FF:FFFFFFFFFFFFFFFFFFFFFFFF:FFFFF:F:FF:FFFFFFFFFFFF      NH:i:0  HI:i:0  AS:i:49 nM:i:4  uT:A:1  xf:i:0  li:i:0  BC:Z:GCCAATCC   QT:Z:FFFF,FFF   CR:Z:TTTGTTGTCTTCGTAT   CY:Z:FFFFFFFFFFFFFFFF   CB:Z:TTTGTTGTCTTCGTAT-1 UR:Z:ACTCCGCTGTCG   UY:Z:FFFFFFFFFFFF   UB:Z:ACTCCGCTGTCG   RG:Z:test:0:1:H2MKKDSXY:4
A00269:304:H2MKKDSXY:4:1242:8223:3865   4   *   0   0   *   *   0   0   CTGAAAAGGGCTTCCAGTCCCGTCACCTTGCCTGCCTGCCTCGGGAGCCAGGGCTGTGCACCTGGCAGTCCCTGCGGTCCCAGATAGCCTGAATCCTGCCCGGAGTGGAAGCTGAAGCCTGCACAGTGTCCACCCTGTTCCCACTCCCAT  FFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFF:FFFFFFFFFFFFFFFFFFFFF:FFFF:FFF:FFFFFFFFFFF:F:FFFF::FFFFFFFFF:FFFFF:FFFFFFFFF:FFFFFFFFFFFFF      NH:i:0  HI:i:0  AS:i:20 nM:i:1  uT:A:1  xf:i:0  li:i:0  BC:Z:AGTTGCAG   QT:Z:FFFFFFFF   CR:Z:TTTGTTGTCTTCGTAT   CY:Z::FFFFFFFFFFFFFFF   CB:Z:TTTGTTGTCTTCGTAT-1 UR:Z:AGTGACCATTCC   UY:Z:FFFFFF:FFFFF   UB:Z:AGTGACCATTCC   RG:Z:test:0:1:H2MKKDSXY:4
A00269:304:H2MKKDSXY:4:2632:25880:13401 4   *   0   0   *   *   0   0   CACACCCCCACGGGAAACAGCAGTGATTAACCTTTAGCAATAAACGAAAGTTTAACTAAGCTATACTAACCCCAGGGTTGGTCAATTTCGTGCCAGCCACCGCGGTCACACGATTAACCCAANTCAATAGAAGCCGGCGTAAAGAGTGTT  FFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFF:FFFF::FFFFFFF:FFFFFFFFFFFFFFFFFFFF:FFFFFFFFFFFFFFFFFFF::FFFFFFFFFFFFFFFFF#FFFFFFFFF:FFFFFFFFFFFFFFFFF      NH:i:0  HI:i:0  AS:i:62 nM:i:9  uT:A:1  xf:i:0  li:i:0  BC:Z:NTGCCGGA   QT:Z:#FFFFFFF   CR:Z:TTTGTTGTCTTCGTAT   CY:Z:FFFFFFFFFFFFFFFF   CB:Z:TTTGTTGTCTTCGTAT-1 UR:Z:CACTTATGTGAT   UY:Z:FFFFFFFFFFFF   UB:Z:CACTTATGTGAT   RG:Z:test:0:1:H2MKKDSXY:4
A00269:304:H2MKKDSXY:4:1573:13621:26522 4   *   0   0   *   *   0   0   AAAAATGCCCTAGCCCACTTCTTACCACAAGGCACACCTACACCCCTTATCCCCATACTAGTTATTATCGAAACCATCAGCCTACTCATTCAACCAATAGCCCTGGCCGTACGCCTAACCGCTAACATTACTGCAGGCCACCTACTCATG  FFFFFFFFFFFFFFFFFF:FFFFFFF,FFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFF:FFFFFF:FFFF:FFFFFFFFFFFF:FFFFFFF:,FF:F:FFFFFFFFFFFFFFFFFFFFFFFFF:FF:FFF,FFFFFF      NH:i:0  HI:i:0  AS:i:19 nM:i:4  uT:A:1  xf:i:0  li:i:0  BC:Z:GCCATTCC   QT:Z:FFFFFFFF   CR:Z:TTTGTTGTCTTCGTAT   CY:Z::FFFFFFFFFFFFFFF   CB:Z:TTTGTTGTCTTCGTAT-1 UR:Z:CTGTTTAATTAG   UY:Z:FFFFFFFFFFFF   UB:Z:CTGTTTAATTAG   RG:Z:test:0:1:H2MKKDSXY:4
A00269:304:H2MKKDSXY:4:1638:20934:35462 4   *   0   0   *   *   0   0   GGCGGATACCATCAAAGCTATTAAGGAAAATCTACCCAAGAAACGCCACACTTTACTGTTCTCAGCCACACAGACCAAAACTCTCAAGGACCAGCCGCGCTGTGGTGGGAAAGACCCTGAGTATGTCTGGGTTCATGAAAAAGCAAAATA  ,FF,FF,:,FF,,:F,FF,:,,:,,,F:FFFFF,:FFFFFF,FFFF:,,,FFF,FFFFFF:FFF:FF,FFFF:FFF::,,FF,F,,::,FFF,,,,,FFF:,,,,F,,::FFF:F,F,FF:FFF,,F,F:FFFFF,,F::FF:FF:FF,F      NH:i:0  HI:i:0  AS:i:69 nM:i:10 uT:A:1  xf:i:0  li:i:0  BC:Z:AGTTGCAG   QT:Z::FFFFFFF   CR:Z:TTTGTTGGCTTCTCAA   CY:Z:,FF:,F,:F:,FFFFF   CB:Z:TTTGTTGTCTTCTCAA-1 UR:Z:AATGGAAAACAG   UY:Z::F,F,FFFFFF,   UB:Z:AATGGAAAACAG   RG:Z:test:0:1:H2MKKDSXY:4
A00269:304:H2MKKDSXY:4:1137:1443:18145  4   *   0   0   *   *   0   0   TCCCAAGAGAGCATCTGTCAGTTGTTGGCAAAATGATAAAGACATTTGAAAATCTGTCTTCTGACAAATAAAAAGCATTTATTTCACTGCACAAAAATAAAAAAAAAAAAAAAAAAAAAAAAAAAAAACGCGGGGTTGGGAAAAAAAACA  FFFF,FFFFFFFFFFFFFF,F:FF,FFFFF,FFFFFF,:FFFF,FFFFF,:FFFFFFFFFFFFFF,,FF:FFFFFF,FFFFFFFF,FFFFF,FFFFFF:FFFFFFFFFFFFFFFFFFFFFFFFFFFF,,F,,F:F,,:F,,,:::F,,,F      NH:i:0  HI:i:0  AS:i:89 nM:i:0  uT:A:1  xf:i:0  li:i:0  BC:Z:TTGCCGGA   QT:Z:FFFFFFFF   CR:Z:TTTGTTGTCTTCTGTA   CY:Z::FFFFFFFFFFFFFFF   CB:Z:TTTGTTGTCTTCTGTA-1 UR:Z:CTCAATCCGACG   UY:Z:FF:FFFFFFFFF   UB:Z:CTCAATCCGACG   RG:Z:test:0:1:H2MKKDSXY:4
A00269:304:H2MKKDSXY:4:1542:18267:19758 4   *   0   0   *   *   0   0   AAAACTAGTTAAGTTTGGGAAAATGAGCTCATGTCTGAAATGAGAAAGTAATTTATAATTTGGTTTATGGGACGGACAGGAAGGAGAAGGAGCACCACTTCTTCCTCAAGTGGAGTGTTTACACCTTGCTGTAACATTTGAACTTTCACA  F:FFFFFFFFFFFFFFFFFFF:FFFFFFFFFFFFFFFFFFFFFFF:FFFF:FFFFFFFFFFFFFFF,FFFFFFFFFF,FFF:FFFFF,F,F:FFFFFFFFFFFFFFFFF:FFFFFFFFFFFFFFFF:FFF:FFFFFFFFFFFFFFFFFFF      NH:i:0  HI:i:0  AS:i:87 nM:i:0  uT:A:1  xf:i:0  li:i:0  BC:Z:AGTTGCAG   QT:Z:FFFFFFFF   CR:Z:TTTGTTGTCTTGAGCA   CY:Z:FFFFFFFFFFFFFFFF   CB:Z:TTTGTTGTCTTGAGCA-1 UR:Z:GTAGGTTTCAAC   UY:Z:FFFFFFFFFFFF   UB:Z:GTAGGTTTCAAC   RG:Z:test:0:1:H2MKKDSXY:4
A00269:304:H2MKKDSXY:4:1278:32570:14278 4   *   0   0   *   *   0   0   ATATAGTGCTTAAGATAGGGCTGAAAATTCTAGAGGAAATTCAGGACAGTCTATCAGAAGTAAAGCGGGGAGAGAGAGTAGGAGCAAGGAGAAACGGTAAGTATACAGGCCTTTCCAAGGGTCTTGAACCCGAGGAAAAGTTAAGGTCAG  FFFFFFFFFFFFFFFFFFFFF:FFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFF:FFFFFFFFFFFFFFFFFFFFFFFF:FF:FF:FFFFFFFFFFFFFFFFFFFFFFFFFFFFF:FFFFFF,::,F::FFF:FF,FFFF:F:FFFF      NH:i:0  HI:i:0  AS:i:148    nM:i:0  uT:A:3  xf:i:0  li:i:0  BC:Z:TTGCCGGA   QT:Z:FFFFFFFF   CR:Z:TTTGTTGTCTTTCTAG   CY:Z:FFFFFFFFFFFFFFFF   CB:Z:TTTGTTGTCTTTCTAG-1 UR:Z:GCTCGACATCTT   UY:Z:FFFFFFFFFFFF   UB:Z:GCTCGACATCTT   RG:Z:test:0:1:H2MKKDSXY:4
A00269:304:H2MKKDSXY:4:2477:17743:31344 4   *   0   0   *   *   0   0   GGAGAGAGAGTAGGAGCAAGGAGAAACGGTAAGTATACAGGCCTTTCCAAGGGTCTTGAACCCGAGGAAAAGTTAAGGTCAGGTAGGAATACCTGGAGAGAGATTAGAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAGTGGGGGAGGA  FFFFFFF:FFFFFFFFFFFFFFFFFFFFFF:FFF:FFF,FFFFFFFFF::FFFFFFFFFFFFFFFFFF,FFFFF,FFFFF:FFF:FFFFFFFFFF:FFF:F:FFFF,FFF,::FFFFFFFF:F:FFF::,FFFFFFFF,,,,,,,,,,,,      NH:i:0  HI:i:0  AS:i:122    nM:i:7  uT:A:3  xf:i:0  li:i:0  BC:Z:AGTTGCAG   QT:Z::FFFFFFF   CR:Z:TTTGTTGTCTTTCTAG   CY:Z:FFFFFFFFFFFFFFFF   CB:Z:TTTGTTGTCTTTCTAG-1 UR:Z:GCTCGACATCTT   UY:Z:FFFFFFFFFFFF   UB:Z:GCTCGACATCTT   RG:Z:test:0:1:H2MKKDSXY:4
[[email protected] bam_files]$ samtools view WT_possorted_genome_bam.bam | tail
A00269:200:HJ7CYDMXX:2:2145:7039:32910  4   *   0   0   *   *   0   0   AAGCAGTGGTATCAACGCAGAGTACATGGGAAAAATTTGTTTCTGTGTGAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA FFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFF:FFFFFFFF:FFFFFFFFFFFFFFF:FFFFF:F:F::F NH:i:0  HI:i:0  AS:i:46 nM:i:1  uT:A:1  xf:i:0  li:i:0  BC:Z:TGTAGAAGQT:Z:FFFFF:FF  CR:Z:TTTGTTGTCTTCGGTC   CY:Z:FFFFFFFFFFFFFFFF   CB:Z:TTTGTTGTCTTCGGTC-1 UR:Z:TTGTCACCATTT   UY:Z:FFFFFFFFFFFF   UB:Z:TTGTCACCATTT       RG:Z:e771c0c8-efbc-45b4-a2f6-c50d2e847e34:0:1:HJ7CYDMXX:2
A00269:200:HJ7CYDMXX:2:2443:9507:13761  4   *   0   0   *   *   0   0   AAGCAGTGGTATCAACGCAGAGTACATGGGATTTGGAATAAAAATTGAATTTCAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA FFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFF:FFFF:FFF,FF:F,:FFF:FF NH:i:0  HI:i:0  AS:i:48 nM:i:3  uT:A:1  xf:i:0  li:i:0  BC:Z:TGTAGAAGQT:Z:FFFFF:FF  CR:Z:TTTGTTGTCTTCGGTC   CY:Z:FFFFFFFFFFFFFFFF   CB:Z:TTTGTTGTCTTCGGTC-1 UR:Z:TTTAAAAACTGC   UY:Z:FFFFFFFFFFFF   UB:Z:TTTAAAAACTGC       RG:Z:e771c0c8-efbc-45b4-a2f6-c50d2e847e34:0:1:HJ7CYDMXX:2
A00269:200:HJ7CYDMXX:1:1239:23375:31234 4   *   0   0   *   *   0   0   CTCGGGGTCGGGGGTGCGTGGCGGGGGCCCGTCCCCCGCCTCCCCTCCGCGCGCCGGGTTTCGCCCCCGCGGCGTCGGGCCCCGCGGACGC FFFFFFFFFFFFFFFFFFFFFFF:FFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFF,FFFFFFFFFFFFFFFFFFFFFF:FFFFFFFFF NH:i:0  HI:i:0  AS:i:54 nM:i:10 uT:A:1  xf:i:0  li:i:0  BC:Z:CTGGACTCQT:Z:FFFFFFFF  CR:Z:TTTGTTGTCTTCGGTC   CY:Z:::FFFFFFF,FFFFFF   CB:Z:TTTGTTGTCTTCGGTC-1 UR:Z:TTTCTGCACATC   UY:Z:F:FFFFFFFFFF   UB:Z:TTTCTGCACATC       RG:Z:e771c0c8-efbc-45b4-a2f6-c50d2e847e34:0:1:HJ7CYDMXX:1
A00269:200:HJ7CYDMXX:1:1181:15736:1626  4   *   0   0   *   *   0   0   AAGCAGTGGTATCAACGCAGAGTACATGGGGTCAAAAGTTTTCCTGCATCTATTAAGATAACCATGGGATTTATGTCCTTGAGATCATTTA FFFFFFFFFFFFFFFFFFFFFFFFFF:FF:F:FFFFFFFFFFFFFF:FFFFFFFFFFFFFFFFF:FFFFF:F,F,FFFFFFF:FFFFFFFF NH:i:0  HI:i:0  AS:i:58 nM:i:1  uT:A:1  xf:i:0  li:i:0  BC:Z:CTGGACTCQT:Z:,F:F:F::  CR:Z:TTAGTTGTCTTCGGTC   CY:Z:F:,:FFF:FFF:::FF   CB:Z:TTTGTTGTCTTCGGTC-1 UR:Z:TTTGGTCTTTTT   UY:Z:F:FF:FFF::F,   UB:Z:TTTGGTCTTTTT       RG:Z:e771c0c8-efbc-45b4-a2f6-c50d2e847e34:0:1:HJ7CYDMXX:1
A00269:200:HJ7CYDMXX:2:1108:14172:13041 4   *   0   0   *   *   0   0   AAGCAGTGGTATCAACGCAGAGTACATGGGGGAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA FFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFF,FFFFFFFFFFFFFFFF:::FF:F:FFFFFFFF NH:i:0  HI:i:0  AS:i:60 nM:i:0  uT:A:3  xf:i:0  li:i:0  BC:Z:AAACCTCAQT:Z:FFFFFFFF  CR:Z:TTTGTTGTCTTCGGTC   CY:Z:FFFFFFFFFFFFFFFF   CB:Z:TTTGTTGTCTTCGGTC-1 UR:Z:TTTTATACCAAT   UY:Z:FFFFFFFFFFFF   UB:Z:TTTTATACCAAT       RG:Z:e771c0c8-efbc-45b4-a2f6-c50d2e847e34:0:1:HJ7CYDMXX:2
A00269:200:HJ7CYDMXX:2:2108:19551:18161 4   *   0   0   *   *   0   0   AAGCAGTGGTATCAACGCAGAGTACATGGGGGAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA FFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFF:FFFFFFFFFFFFFFFFFFF:F,FF,FFFFFFFFF:F NH:i:0  HI:i:0  AS:i:60 nM:i:0  uT:A:3  xf:i:0  li:i:0  BC:Z:AAACCTCAQT:Z:FFFFFFFF  CR:Z:TTTGTTGTCTTCGGTC   CY:Z:FFFFFFFFFFFFFFFF   CB:Z:TTTGTTGTCTTCGGTC-1 UR:Z:TTTTATACCAAT   UY:Z:FFFFFFFFFFFF   UB:Z:TTTTATACCAAT       RG:Z:e771c0c8-efbc-45b4-a2f6-c50d2e847e34:0:1:HJ7CYDMXX:2
A00269:200:HJ7CYDMXX:2:2173:30553:22717 4   *   0   0   *   *   0   0   AAGCAGTGGTATCAACGCAGAGTACATGGGGTCCCAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA FFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFF:FFFFFFFFFFFFFF,FFFF,FFF:FFFF NH:i:0  HI:i:0  AS:i:57 nM:i:0  uT:A:1  xf:i:0  li:i:0  BC:Z:CTGGACTCQT:Z:F:FFFFFF  CR:Z:TTTGTTGTCTTCGGTC   CY:Z:FFFFFFFFFFFFFFFF   CB:Z:TTTGTTGTCTTCGGTC-1 UR:Z:TTTTCTACTTGG   UY:Z:FFFFFFFFFFFF   UB:Z:TTTTCTACTTGG       RG:Z:e771c0c8-efbc-45b4-a2f6-c50d2e847e34:0:1:HJ7CYDMXX:2
A00269:200:HJ7CYDMXX:2:2173:30798:22764 4   *   0   0   *   *   0   0   AAGCAGTGGTATCAACGCAGAGTACATGGGGTCCCAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA FFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFF:FFFFFFFFFF:FFFFFFFFFFFF:FFFFF:,FFFFF:FFFFFFF NH:i:0  HI:i:0  AS:i:57 nM:i:0  uT:A:1  xf:i:0  li:i:0  BC:Z:CTGGACTCQT:Z:FFFFFFFF  CR:Z:TTTGTTGTCTTCGGTC   CY:Z:FFFFFFFFFFFFFFFF   CB:Z:TTTGTTGTCTTCGGTC-1 UR:Z:TTTTCTACTTGG   UY:Z:FFFFFFFFFFFF   UB:Z:TTTTCTACTTGG       RG:Z:e771c0c8-efbc-45b4-a2f6-c50d2e847e34:0:1:HJ7CYDMXX:2
A00269:200:HJ7CYDMXX:1:2435:31403:9502  4   *   0   0   *   *   0   0   GCTGGGACAGCCCCGCTCCCAGATGTAAAGAACGCGACTTCCACAAACCTGGATTTTTTATGTACAACCCTGACCGTGACCGTTTGCTATA :,FFFFF:F:FFFFFFFFFFF:FF,FFFFFFFFFFFFFFFFFFFFFFFFFFFFF,FFF:FFF,FFFFFFFFFFF:FFFFFFFFFFFFFFFF NH:i:0  HI:i:0  AS:i:20 nM:i:0  uT:A:1  xf:i:0  li:i:0  BC:Z:GCCTTGGTQT:Z:FFFF:FFF  CR:Z:TTTGTTGTCTTGGATG   CY:Z:FFFFF:FFFFFF::FF   CB:Z:TTTGTTGTCTTGGATG-1 UR:Z:AATTCACGTACT   UY:Z:FFFFFFFFFFFF   UB:Z:AATTCACGTACT       RG:Z:e771c0c8-efbc-45b4-a2f6-c50d2e847e34:0:1:HJ7CYDMXX:1
A00269:200:HJ7CYDMXX:2:2457:17743:34507 4   *   0   0   *   *   0   0   AAGCAGTGGTATCATCGCAGATTACATGGGCAATTTCACTAATTCCAATACTTATTATGATTGAAACAATTAGACTATGTCTTCAACCAAT ::FFFFFFFFFFFF,FFFFFF,,FFFF,F:FFFF:F:FFFFFFFF,,FFFF:F,F,FF:FFF,F,FFFFFFFF,FFFF,:FF,FFFFF,F: NH:i:0  HI:i:0  AS:i:52 nM:i:4  uT:A:1  xf:i:0  li:i:0  BC:Z:GCCATGGTQT:Z:,,F,:F,:  CR:Z:TTTGTTGTCTTTGGAG   CY:Z:FF:F:FFF,F,,,FF:   CB:Z:TTTGTTGTCTTTGGAG-1 UR:Z:CATGTTCCGTCC   UY:Z:FFFF:,,::FFF   UB:Z:CATGTTCCGTCC       RG:Z:e771c0c8-efbc-45b4-a2f6-c50d2e847e34:0:1:HJ7CYDMXX:2

Could you please help me with this issue?

Thanks in advance!
Yi



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