Several lysis buffers can isolate nuclei from fresh or frozen tissues. However, I am not sure the ratio of lysis buffer to tissue. Some tissues with muscle and smooth muscle have more debris after lysis. Does it mean that I should add more lysis Buffer or prolong the time? Do not have enough lysis buffer will cause rna degation?
Tania L Gonzalez
The debris comes from 2 sources: your storage of the frozen tissue (more buffer or water will cause ice crystals to break cells and increase debris) and over-lysing the sample during processing. I would focus on improving your sample storage if you are getting new samples or optimizing your lysis time. Sorting the nuclei may be possible also if your protocol separates the isolation and permeabilization steps.
1 votes
1 thanks
More Hyde Chen's questions See All
Similar questions and discussions