10x Single Cell Multiome Nuclei Isolation and Library Preparation Protocol v1

10x single-cell multiome nuclei isolation and library preparation protocol for HuBMAP colon project.

Protocol for nuclei isolation from fresh and frozen tissues using Salty-Ez10 or Salty-Ez50 buffer: compatible with snRNA-Seq and Multiome workflows from 10x Genomics v2

This is a protocol in development, which means it has not yet tested/challenged with multiple samples. So, please make sure you take it for a test drive before committing to it. Once you do please share your experience with me via email, Twitter or as a comment. In this version I labelled DTT as optional since after further testing I have not seen any difference with or without in snRNA-Seq workflow. Also, I recommend WRB1 for snRNA-Seq workflow only (although still works fine for Multiome) and WRB2 for both snRNA-Seq and Multiome workflows. Since WRB2 is 10x Genomics' recommendation for Multiome I have adopted this for this workflow. I include a discussion about cycling during cDNA amp. SaltyEz50 option: this has been particularly useful for samples where lysis in SaltyEz10 showed to be suboptimal. In my hands, those were breast, liver and pancreas.