We are trying to isolate nuclei from mouse skeletal muscle but at the end of the process, the clumping of nuclei was still present in the slide. We were not getting good-quality of nuclei for library preparation and further genomics study.
As a resuspended buffer, I used RNAse inhibitor, 2% BSA and 1X PBS buffer. So, I would like to know how to minimize/remove these clumping nuclei and get better nuclei quality at the final stage after the resuspension process. Please anyone can help me how to solve this problem?
Thanks in advance
Traditionally, Non-ionic detergent is used for isolating nuclei from tissues. It is the detergent that causes nuclear clamping. You can try detergent-free method or a detergent -containing kit but is able to generating single nuclei.
https://inventbiotech.com/products/minute-detergent-free-nuclei-isolation-kit?_pos=1&_sid=48dff6ceb&_ss=r
https://inventbiotech.com/products/minute%E2%84%A2-single-nucleus-isolation-kit-for-tissues-cells-20-preps?_pos=2&_sid=48dff6ceb&_ss=r
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