<div dir="ltr"><div>I am new, please help, I got InvalidFASTQFileFormat error, the sequence looks fine, even after I remove that sequence, it still shows the error, and the error always show on line 16181.</div><div><br></div>

<div>[17:27:40]sw4ed@hpcserver:~/Meta/SoilMeta&gt; load-into-counting.py -T 4 -k 20  -N 4 -x 4e9 Sample_2024-1F_1.kh Sample_2024-1F_1.trimmed.fastq</div><div><br></div><div>PARAMETERS:</div><div> - kmer size =    20            (-k)</div>

<div> - n hashes =     4             (-N)</div><div> - min hashsize = 4e+09         (-x)</div><div><br></div><div>Estimated memory usage is 1.6e+10 bytes (n_hashes x min_hashsize)</div><div>--------</div><div>Saving hashtable to Sample_2024-1F_1.kh</div>

<div>Loading kmers from sequences in [&#39;Sample_2024-1F_1.trimmed.fastq&#39;]</div><div>making hashtable</div><div>consuming input Sample_2024-1F_1.trimmed.fastq</div><div>terminate called after throwing an instance of &#39;khmer::read_parsers::InvalidFASTQFileFormat&#39;</div>

<div>  what():  InvalidFASTQFileFormat: illegal sequence letters: @HISEQ700708:147:D278GACXX:2:1101:6816:3815 2:N:0:ATCACG</div><div>Aborted (core dumped)</div><div><br></div><div><div>[17:44:12]sw4ed@hpcserver:~/Meta/SoilMeta&gt; sed -n &#39;16181,16188p&#39; Sample_2024-1F_1.trimmed.fastq</div>

<div>@HISEQ700708:147:D278GACXX:2:1101:6816:3815 2:N:0:ATCACG</div><div>ATTGTCTGCGCGTTACGATATTATCAAGAATCGCGACTGGTTATGGTCTCTTACTGCTACAACACTGAACACTAAGACCAAATATGCTAACATTGGCAAC</div><div>+</div><div>CCCFFFFFHHHHHIJJJJIJJJJJJJJJJJJJJJJJJJJJIJIIIJIHHHHHFFFFFFFFEEEDDDDDDDDDDCDDDDDDDDDDEEDEDDDDDDDDDDDD</div>

<div>@HISEQ700708:147:D278GACXX:2:1101:6919:3932 1:N:0:ATCACG</div><div>TATTCAGGAAAACCTGCCGCAGACGCTTGGGGTCGCAGGAGATTTCCGGAATGTCTTCGTCATTGTCGATATAGTCCAGTCGAATACCGTCCTGGGCCAG</div><div>+</div><div>@C@FFFDFFHGFHIGIIJIIIJJJJGIJIJJIFEFHIIGGDHFHHHHFF&lt;ACAEDDCDD=?A?DD@CBDDBDDCC@CDCDD@B?C@CCB&lt;@DD&gt;C?BDDB</div>

</div><div><br></div><div><br></div>-- <br><div dir="ltr">Shaolin Wang, Ph.D<br>Research Scientist<br>Department of Psychiatry &amp; Neurobiology Science<br>University of Virginia<br>1670 Discovery Drive, Suite 110<br>Charlottesville, VA 22911<br>

<font face="Arial, sans-serif">
</font>Phone: <a href="tel:434-982-0243" value="+14349820243" target="_blank">434-982-0243</a><br>Fax:<a href="tel:434-973-7031" value="+14349737031" target="_blank">434-973-7031</a><br>E-mail: <span><span><a href="mailto:sw4ed@eservices.virginia.edu" target="_blank">swang@virginia.edu</a></span></span><div style="display:inline">
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