We are trying to demultiplex our samples using Cite-seq but we are getting low overlap between cell barcodes and hashtag barcodes. We used the same pipeline on our previous experiment and didn't have this issue. Here is a summary of our experiment :
Experimental library summary:
• Nuclei samples
• Nuclei HTO staining
• Added HTO cDNA PCR additive primer (V1)
• Used 10x dual index plate TT for both HTO library and transcriptome
• Not great overlap at demultiplexing step post sequencing
We are trying to demultiplex our samples using Cite-seq but we are getting low overlap between cell barcodes and hashtag barcodes. We used the same pipeline on our previous experiment and didn't have this issue. Here is a summary of our experiment :
Experimental library summary: • Nuclei samples • Nuclei HTO staining • Added HTO cDNA PCR additive primer (V1) • Used 10x dual index plate TT for both HTO library and transcriptome • Not great overlap at demultiplexing step post sequencing
Could you please help us to resolve this issue?