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Size of the small pieces of the mince process?

Ruuheena Peng Jun 24, 2022

Protocol: Generation and Screening of a Non-typeable Haemophilus influenzae Tn-seq Mutant Library

Dear authors,

In the mitochondrial extraction process, you mentioned that muscles should be minced into small pieces. I am wondering to what extent we need to mince them. I am also curious whether it is possible to extract mitochondria specifically from muscle satellite cells usin Read more Read less more less

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Alexandre Prola Answered Jun 27, 2022

Alexandre Prola

Dear Ruuheena Peng,

Thank you for your interest.

We typically mince muscles with small scissors for approximately 2 min on ice, in order to obtain very small pieces of muscle. This allows optimal action of trypsin in the following step.

Regarding the extraction of mitochondria from satellite cells, we never tried but is should work with minor adjustments, I guess you can use the procotol described for MEF in the following paper:

https://pubmed.ncbi.nlm.nih.gov/17406588/

Good luck!

Alexandre

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