Hi all,
I am starting this thread to connect and share resources/knowledge on sample prep protocols for extracting nuclei to performing snRNA seq experiments.
The main issue with this is that there are too many published protocols using different methods and it is hard to understand the rational behind each reagent and process so that one can optimize their protocol to make it suit their needs.
Main issues I faced are
1) increased cell debris/myelin
2) nuclei clumping
3) overlysis (clumping and unhealthy nuclei)/underlysis (more cells/nuclei ratio)
I am hoping to have a focussed discussion on this and willing to share my experiences with different protocols I used.
I am wondering if someone used the 10x kit or Miltenyi protocol with success for the brain tissues.
Thanks in advance.