unsupported operand type(s) for +=: 'int' and 'NoneType' in 2024

[Ohmori-Insilico]

We are having this problem using the SAM file to identify the off-target with Py3.

(base) user@ohmori-lab-ubuntu:~/Desktop/Guideseq 2024/guideseq/guideseq$ guideseq.py identify \

--aligned Data/aligned/VEGFA1_Tsai.sam \ --genome Homo_sapiens_GRCh38ssembly.fa \ --outfolder "/home/user/Desktop/Guideseq 2024/guideseq/guideseq/output" \ --target_sequence GGGTGGGGGGAGTTTGCTCCNGG \ --description "VEGFA1_site1" [04/02 07:46:48AM][INFO][guideseq] Identifying offtarget sites... [04/02 07:46:48AM][INFO][identifyOfftargetSites] Processing SAM file /home/user/Desktop/Guideseq 2024/guideseq/guideseq/output/aligned/VEGFA1_Tsai.sam [04/02 07:47:01AM][ERROR][guideseq] Error identifying offtarget sites. [04/02 07:47:01AM][ERROR][guideseq] Traceback (most recent call last): File "/home/user/miniconda3/bin/guideseq.py", line 239, in identifyOfftargetSites identifyOfftargetSites.analyze(samfile, self.reference_genome, self.identified[sample], annotations, File "/home/user/miniconda3/bin/identifyOfftargetSites.py", line 314, in analyze chromosome_position.addPositionBarcode(chromosome, read_position, strand, barcode, primer, count) File "/home/user/miniconda3/bin/identifyOfftargetSites.py", line 60, in addPositionBarcode self.chromosome_barcode_dict[chromosome][position][strand][barcode] += count TypeError: unsupported operand type(s) for +=: 'int' and 'NoneType'

Samfile size: 7GB. I used UMI, consolidation and aligned data. But problem is still persisted. Test with EMX1 data from Guideseq insruction, it is working. Is file size matter ?

Is there any solution to complete this task ?

Thanks a lot.

Neme.