Cell Culture: cell-line misidentification

1. Ranked causes

   1. Cross-contaminated or swapped vial (wrong line in the flask)STR / authentication missing or overdue; marker panel mismatches certificate.
   2. Over-passaged stock drifted from the authenticated early-passage phenotypeEarly-passage authenticated vial still matches; late passage does not.
   3. Vendor or collaborator stock was already misidentified upstreamAuthentication fails on the unopened vendor vial as well.
   4. Labeling or cryostock inventory mix-up at thawBox map or vial label disagrees with the expected RRID / catalog number.

2. Recovery steps

   1. Authenticate the cell line (STR / species PCR) and record lastAuthenticatedAt on Lab IndexDirectly tests the leading material cause and updates Lab Index dates.
   2. Quarantine the suspect flask; thaw an authenticated early-passage vial in parallelStops spreading a wrong line while confirmation runs.
   3. Reconcile cryostock box map against vial labels and RRIDs before the next thawAddresses inventory mix-ups that present as “wrong phenotype”.
   4. If the unopened vendor vial also fails authentication, contact the vendor and stop using the lotUpstream misidentification cannot be fixed by lab technique alone.

3. Controls first

   • Did a known authenticated positive-control line pass the same STR / species assay?If no: Assay or kit failure — fix the authentication assay before discarding stocks.
   • Does an unopened vendor vial of the same lot also fail authentication?If yes: Upstream stock issue — quarantine the lot and notify the vendor.

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