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kmnip avatar kmnip commented on July 23, 2024

Hello @mhfk ,

The assembler crashed because the abyss-map aligner expects the mates to be in opposite orientations, i.e.
----> <----
So, you should use the original reads as input to Trans-ABySS.

Different in the log refers to the number of read pairs that are aligned to different unitigs.
The read orientations (i.e. FF, RF, and FR) are tallied based on the mates aligned to single unitigs. I am actually quite surprised that there are only 112 reads for this.

Ka Ming

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mhfk avatar mhfk commented on July 23, 2024

Hi @kmnip ,
Apologies for the late follow-up. I have tried to run the alignment using the original reads and the same issue persisted. My workstation froze so I couldn't get the screenshot but the same error message prompted. I wonder if the assembler identified my reads as unstranded and couldn't proceed with assembly.

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kmnip avatar kmnip commented on July 23, 2024

Can you verify that the read names have the suffixes /1 and /2 within your read files?

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mhfk avatar mhfk commented on July 23, 2024

I think the file names might be the issue. The names of the files were :
Paired-end

91_1.paired.fq.gz & 91_2.paired.fq.gz
92_1.paired.fq.gz & 92_2.paired.fq.gz
93_1.paired.fq.gz & 93_2.paired.fq.gz
94_un-conc-mate.1.paired.fq.gz & 94_un-conc-mate.2.paired.fq.gz
95_un-conc-mate.1.paired.fq.gz & 95_un-conc-mate.2.paired.fq.gz
96_un-conc-mate.1.paired.fq.gz & 96_un-conc-mate.2.paired.fq.gz \

Single-end

91_1.unpaired.fq.gz & 91_2.unpaired.fq.gz & 92_1.unpaired.fq.gz & 92_2.unpaired.fq.gz & 93_1.unpaired.fq.gz & 93_2.unpaired.fq.gz
94_un-conc-mate.1.unpaired.fq.gz & 94_un-conc-mate.2.unpaired.fq.gz
95_un-conc-mate.1.unpaired.fq.gz & 95_un-conc-mate.2.unpaired.fq.gz & un-singleton.trimmed.fq.gz
96_un-conc-mate.1.unpaired.fq.gz & 96_un-conc-mate.1.unpaired.fq.gz & un-singleton.trimmed.fq.gz

I am changing the file names to names with strand information at the back (*_1.fq.gz or *_2.fq.gz) instead of having them in the middle as seen above. Will let you know if this works!

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mhfk avatar mhfk commented on July 23, 2024

@kmnip
The error persisted with similar error prompt. Perhaps there is something wrong with my command. I have both PE and SE reads to be assembled. SE reads belong to SE library and residual data from Trimmomatic. Here are the commands :

`transabyss --SS --threads 16 --pe \

./axenic/SRR8432491/91.paired_1.fq.gz ./axenic/SRR8432491/91.paired_2.fq.gz
./axenic/SRR8432492/92.paired_1.fq.gz ./axenic/SRR8432492/92.paired_2.fq.gz
./axenic/SRR8432493/93.paired_1.fq.gz ./axenic/SRR8432493/93.paired_2.fq.gz
./inplanta/SRR8432494/94_un-conc-mate.paired_1.fq.gz ./inplanta/SRR8432494/94_un-conc-mate.paired_2.fq.gz
./inplanta/SRR8432495/95_un-conc-mate.paired_1.fq.gz ./inplanta/SRR8432495/95_un-conc-mate.paired_2.fq.gz
./inplanta/SRR8432496/96_un-conc-mate.paired_1.fq.gz ./inplanta/SRR8432496/96_un-conc-mate.paired_2.fq.gz
--se
./axenic/SRR8432491/91.unpaired_1.fq.gz ./axenic/SRR8432491/91.unpaired_2.fq.gz ./axenic/SRR8432492/92.unpaired_1.fq.gz ./axenic/SRR8432492/92.unpaired_2.fq.gz ./axenic/SRR8432493/93.unpaired_1.fq.gz ./axenic/SRR8432493/93.unpaired_2.fq.gz
./inplanta/SRR8432494/94_un-conc-mate.unpaired_1.fq.gz ./inplanta/SRR8432494/94_un-conc-mate.unpaired_2.fq.gz
./inplanta/SRR8432495/95_un-conc-mate.unpaired_1.fq.gz ./inplanta/SRR8432495/95_un-conc-mate.unpaired_2.fq.gz ./inplanta/SRR8432495/un-singleton.trimmed.fq.gz
./inplanta/SRR8432496/96_un-conc-mate.unpaired_1.fq.gz ./inplanta/SRR8432496/96_un-conc-mate.unpaired_2.fq.gz ./inplanta/SRR8432496/un-singleton.trimmed.fq.gz `

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