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The following example is for planning RNP complexes for 1 response. Change accordingly dependant on the quantity of reactions needed.
Movement cytometry can be employed to measure the performance of differentiation protocols While using the STEMdiff™ system, by checking out particular marker expression at Each and every stage with the differentiation procedure. It can even be used for other apps, including cell sorting, immunophenotyping, and purity assessment.
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Set the “unstained” tube apart. For that “viability dye” sample, centrifuge the tube at three hundred x g
Consider the standing by checking a droplet and halting the dissociation when all around eighty% on the cell suspension is solitary-celled. Prevent about-digestion.
If bigger clumps remain visible in the answer, return the tube to 37°C for an additional 2 minutes and repeat the procedure right up until organoids have completely broken into one cells.
When organoids are launched from Matrigel® and floating in the harvesting Option, transfer everything right into a clean conical tube. Allow organoids settle by gravity or by centrifuging the tube for five minutes at two hundred x g
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Notes: Use pipette ideas to Carefully scratch The underside of each and every well to be able to raise the monolayer off, then dissociate the cell clumps into one-cell suspension by pipetting up and down five - ten situations.
That is an end-point protocol designed Stem cell to dissociate PSC-derived kidney organoids into a single-cell suspension and fixation for downstream move cytometry Assessment.
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