Hi, I have run harmony (and all pre-normalizing steps) and when I get to RunUmap, my umap is essentially split by seq type. I have ran this data before in different subsets and the flex and sc data has clustered well together. There are usually some clusters unique to seq type but I found they were real. Here, however the same celltypes are separated by seq type as you can see. I am wondering if it has to do with alignment? Any advice would be appreciated. To merge these two seq types I create a seurat object for both and merge/join them. I have tried normalizing before and after this step as well. Not sure if there has been updates to packages causing these problems. Like I said this has worked before- so I am lost at why it won't now. Thank you! https://preview.redd.it/3zhu511zap3h1.png?width=700&format=png&auto=webp&s=3751f146f8e51dea6ad8d24c98c7b9850b9f35f2 submitted by /u/liltthebasedg0d [link] [comments]