Single Cell Extraction

Tumor dissociation protocol

Prepare digestion medium before going for tumor collection: 20 ml RPMI containing 0.05 mg/ml TM liberase and 0.02 mg/ml DNase I for each tumor. TM stock: 5 mg/ml (200 µl); DNase I stock: 10mg/ml (40 µl).

Perform mechanical dissociation

Perform enzymatic dissociation

Remove RBC and quantify viable cells

TM liberase (Roche)

10x Genomics – Removal of Dead Cells from Single Cell Suspension for Single Cell RNA Sequencing

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