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Hello,
I am running cellranger_workflow with v8.0.1 with SC3Pv4 chemistry where we used TotalSeqB antibodies for sample hashing as well as for CITE-seq. The workflow runs without any errors, and I have over 97% of my reads with valid cell barcodes. However, I noticed that in the adt count log file, it is detecting TotalSeq-C instead of TotalSeq-B, with about 30% of indices being ambiguous. When I then try to find matching cell barcodes between my matched gene and adt count matrix, there is only about a 1% overlap between the two. I also checked if there was any sample mismatch, but I get very low overlap across all combinations of my adt and gene libraries.
I was wondering then if the TotalSeq mismatch may contribute to why I am seeing a very low overlap between my gene expression and antibody libraries? I have ran this workflow before with cellranger v7.0 using v3.1 chemistry and it has worked great, so any help here would be appreciated!
Hello,
I am running cellranger_workflow with v8.0.1 with SC3Pv4 chemistry where we used TotalSeqB antibodies for sample hashing as well as for CITE-seq. The workflow runs without any errors, and I have over 97% of my reads with valid cell barcodes. However, I noticed that in the adt count log file, it is detecting TotalSeq-C instead of TotalSeq-B, with about 30% of indices being ambiguous. When I then try to find matching cell barcodes between my matched gene and adt count matrix, there is only about a 1% overlap between the two. I also checked if there was any sample mismatch, but I get very low overlap across all combinations of my adt and gene libraries.
I was wondering then if the TotalSeq mismatch may contribute to why I am seeing a very low overlap between my gene expression and antibody libraries? I have ran this workflow before with cellranger v7.0 using v3.1 chemistry and it has worked great, so any help here would be appreciated!
Thanks,
Manny
Below I have attached the adt log.txt file, my sample sheet, feature barcode file, and the adt.report.txt file:
adtlog.txt
IFNB_B2_adt.report.txt
sample_sheet.csv
feature_barcode.csv
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