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Will the CometAssay® work with solid tissue? If so how?

PostPosted: Mon 16 Feb 2015 15:33
by Interchim
Will the CometAssay® (4250-050-K) work with solid tissue? If so how?
FAQ from Trevigen

Carefully prepared cell suspensions from tissues can be used with the CometAssay®.

Protocol Guideline

1. Place a small piece of tissue in 1-2 ml of cold HBSS containing 20 mM EDTA.
2. Mince tissue into fine pieces and allow settling.
3. Remove 5-10 µl of the supernatant (cell suspension) and combine with 75 µl molten LMAgarose.

To ensure the cell concentration and dissociation are sufficient, a small aliquot can be diluted in PBS (instead of LMAgarose) and spotted onto a microscope slide.

Another option is to digest the tissue for 15 minutes with trypsin, wash with HBSS or cold PBS containing serum to inactivate the enzyme and then mechanically dissociate using syringe and needle.