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Error in running kissplice2reftranscriptome

Dear all ! I've successfully run the KisSplice2RefTranscriptome workflow according to the documentation, I got 30 reads.fq and finish Transdecoder with a reference fasta file (Using Pacbio Seq to sequence and create)like this :

>Gene.103793
ATGTGTGACGACGAGGAAAGCACTGCCCTTGTGTGTGACAATGGATCAGGACTGTGTAAG
GCAGGCTTCGCCGGGGATGATGCTCCCAGAGCGGTGTTCCCCTCCATAGTGGGTCGTCCC
AGACACCAGGGAGTGATGGTGGGAATGGGGCAGAAAGATAGTTATGTTGGCGATGAGGCG
CAAAGCAAAAGGGGAATCCTGACCCTCAAGTACCCTATCGAGCACGGCATCATCACAAAC
TGGGACGACATGGAAAAAATCTGGCATCACTCTTTCTACAATGAGCTCCGTGTGGCTCCA
GAGGAGCACCCCACTCTGCTGACAGAAGCCCCTCTCAATCCCAAAGCCAACAGGGAAAAG
ATGACCCAGATCATGTTTGAGACCTTCAATGTTCCAGCCATGTATGTAGCTATCCAAGCG
GTGCTGTCCCTGTATGCTTCTGGACGTACAACAGGCATTGTCCTCGACTCTGGTGATGGT
GTCACCCACAACGTGCCCATTTATGAGGGCTACGCTCTTCCGCATGCCATTATGCGTTTG
GACTTGGCCGGCCGTGATCTCACTGACTACCTCATGAAGATCCTAACTGAACGAGGCTAC
TCATTCGTCACCACTGCTGAAAGAGAAATTGTCCGAGACATCAAAGAGAAGTTGTGCTAT
GTGGCGCTGGATTTTGAGAATGAGATGGCCACTGCAGCCTCTTCCTCCTCCCTGGAGAAG
AGCTATGAGCTTCCCGATGGACAGGTCATCACCATTGGCAATGAGAGGTTCCGCTGCCCT
GAGACACTCTTCCAGCCATCCTTCATTGGAATGGAATCTGCTGGCATCCATGAAACCACT
TACAACAGCATCATGAAATGCGACATTGATATCCGTAAGGACCTGTATGCCAACAACGTG
TTGTCTGGTGGTACCACCATGTACCCTGGCATTGCAGACAGGATGCAGAAGGAGATCACT
GCTTTGGCCCCCAGCACCATGAAGATCAAGATCATCGCCCCACCTGAGCGCAAGTACTCA
GTATGGATTGGCGGCTCCATCCTGGCCTCCCTCTCCACCTTCCAGCAGATGTGGATAAGC
AAGCAGGAGTATGACGAGGCCGGGCCATCCATTGTGCATCGCAAGTGCTTCTAA

the error occur in this step:

python3 kissplice2reftranscriptome -b NJLFY.transdecoder.cds.fa.bed -k results_coherents_type_2.fa -t type_coherent_2.psl

the problem is:

Opening all files and formatting tables...
Collecting data...
Traceback (most recent call last):
  File "kissplice2reftranscriptome", line 577, in <module>
    main()
  File "kissplice2reftranscriptome", line 443, in main
    blatbd.lowerPath, kSize)
  File "/home/xuweitong/software/kisSplice/kissplice2reftranscriptome-1.3.3/kissplice2reftranscriptomelib/dataAnalysis/snpPositionFinder.py", line 54, in getMultipleSnpPositionInBubbleWithBlat
    if upperRowNucleotide[i] != lowerRowNucleotide[i]:
IndexError: string index out of range

the file results_coherents_type_2.fa is :

>bcc_92143|Cycle_0|Type_2|upper_path_length_91|C1_0|C2_0|C3_0|C4_0|C5_0|C6_0|C7_0|C8_0|C9_2|C10_0|C11_0|C12_0|C13_0|C14_0|C15_0|C16_0|C17_0|C18_0|C19_0|C20_0|C21_0|C22_0|C23_0|C24_0|C25_0|C26_0|C27_0|C28_0|C29_0|C30_0|rank_1.00000
AACTGTATTTTGAAATTCAGAAATAGACAACTTATCACTTGTACAAACTAATCACTTGCACAAACTTATCACTTGTACAAACTTATCACTT
>bcc_92143|Cycle_0|Type_2|lower_path_length_74|C1_1|C2_1|C3_0|C4_0|C5_1|C6_0|C7_0|C8_0|C9_0|C10_0|C11_0|C12_0|C13_0|C14_0|C15_0|C16_0|C17_0|C18_0|C19_0|C20_0|C21_0|C22_0|C23_3|C24_3|C25_0|C26_0|C27_0|C28_0|C29_0|C30_0|rank_1.00000
AACTGTATTTTGAAATTCAGAAATAGACAACTTATCACTTGCACAAACTTATCACTTGTACAAACTTATCACTT

and the file type_coherent_2.psl is :
psLayout version 3

match   mis-    rep.    N's Q gap   Q gap   T gap   T gap   strand  Q           Q       Q       Q   T           T       T       T   block   blockSizes  qStarts  tStarts
        match   match       count   bases   count   bases           name        size    start   end name        size    start   end count
---------------------------------------------------------------------------------------------------------------------------------------------------------------
34  0   0   0   0   0   1   30  +   bcc_75492|Cycle_50|Type_2|upper_path_length_86|C1_0|C2_0|C3_0|C4_0|C5_0|C6_0|C7_0|C8_0|C9_0|C10_0|C11_0|C12_0|C13_0|C14_0|C15_0|C16_0|C17_0|C18_0|C19_0|C20_0|C21_0|C22_0|C23_0|C24_0|C25_4|C26_0|C27_0|C28_0|C29_0|C30_0|rank_1.00000  86  52  86  Gene.94817  1809    1429    1493    2   17,17,  52,69,  1429,1476,
39  0   0   0   0   0   2   71  +   bcc_75492|Cycle_50|Type_2|upper_path_length_86|C1_0|C2_0|C3_0|C4_0|C5_0|C6_0|C7_0|C8_0|C9_0|C10_0|C11_0|C12_0|C13_0|C14_0|C15_0|C16_0|C17_0|C18_0|C19_0|C20_0|C21_0|C22_0|C23_0|C24_0|C25_4|C26_0|C27_0|C28_0|C29_0|C30_0|rank_1.00000  86  35  74  Gene.80066  1245    1085    1195    3   13,12,14,   35,48,60,   1085,1113,1181,

Appreciate for any info! Thanks in advance!

snp kissplice rna-seq

1 answer

Dear user,

KisSplice outputs several files. The one for SNPs is "results_coherents_type_0.fa" I think that the bug comes from the fact that you are using "results_coherents_type_2.fa", which corresponds to inexact tandem repeats. KisSplice2RefTranscriptome only handles SNPs.

Best,

Vincent

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