BCPAP (65 (link)) and K1 (66 ) cells were purchased from ATCC in 2014 and maintained in RPMI supplemented with 10% fetal bovine serum, 1% penicillin, and 1% streptomycin. The identity of both cell lines was confirmed by DNA profiling of polymorphic short tandem repeat (STR) markers through the human cell line authentication analysis service at the Duke University DNA Analysis Facility. The resultant STR makers, assess by GenePrint 10 kit (Promega), were compared to those available for BCPAP (CVCL_0153) and K1 (CVCL_2537) cells lines through Cellosaurus (65 (link)) on February 2018. Both cell lines were also confirmed to be free of mycoplasma infection, as assessed by the Duke Cell Culture Facility using MycoAlert PLUS test (Lonza) on January 2018. Both cell lines were used within 5 passages of being thawed. BCPAP cell lines were engineered to express ERK2GOF by stable infection using established methods (67 (link)) of a retrovirus derived from the plasmid pBABEpuro-HA-ERK2GOF encoding the ERK2R67S, D321N mutant form of ERK2, termed ERKGOF (40 (link)).
Protocol detail
Cell Line Authentication and Modification
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Cell culture
Cell line authentication
Cell lines
Cells
Dna markers
Erk2
Fetal bovine serum
Human
Infection
Mycoplasma infection
Penicillin
Plasmid
Polymorphic
Promega
Retrovirus
Short tandem repeat
Streptomycin
Corresponding Organization : Duke Medical Center
Other organizations : Clinical Research Institute
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Variable analysis
independent variables
- ERK2^GOF expression (engineered BCPAP cell lines)
- Not explicitly mentioned
- Maintenance of both BCPAP and K1 cell lines in RPMI supplemented with 10% fetal bovine serum, 1% penicillin, and 1% streptomycin
- Confirmation of cell line identity through DNA profiling of polymorphic short tandem repeat (STR) markers
- Confirmation of absence of mycoplasma infection
- Usage of cell lines within 5 passages of being thawed
- Positive control: BCPAP (CVCL_0153) and K1 (CVCL_2537) cell lines as reference standards for STR marker comparison
- Negative control: Not explicitly mentioned
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