mcbStability testing of genetically engineered prokaryotic or eukaryotic cells is essential to assure the quality of production cell banks. ACGT offers genetic stability testing to characterize Master Cell Banks (MCB), Working Cell Banks (WCB), and End of Production Cells (EPC) from bacterial, yeast, insect, and vertebrate cell cultures.

Our analysis begins with the isolation of a sufficient number of clones to adequately sample the population of cells in the culture. Copy number determination, NGS analysis, and sequencing are performed on DNA prepared from the clones. We provide sequence verification by direct analysis of RT-PCR fragments targeting the appropriate mRNA for the MCB as well as the WCB and EPC.  NGS and quantitative PCR analysis are performed to confirm the stability and chromosome location of the inserted expression cassettes.

Service Description:
  • Both absolute quantitation (AQ) and relative quantitation (RQ) RT-qPCR
  • DNA/RNA extraction from MCBs, and WCBs, and EOPs
  • Transgene insertion copy number analyzed by qPCR and/or NGS
  • Phage detection and residual DNA analyzed by qPCR and/or NGS
  • Structural analysis (insertion sites/restriction analysis) by Southern blot
  • Transcriptome analysis by RT-PCR and DNA sequencing
  • RT-qPCR for expression analysis
mcb2Data Archival at the Conclusion of the Study (per Title 21 CFR):
  • Original test articles for a minimum of 2 years
  • Hard copy of the final protocol, Final Report and any amendments
  • Laboratory notebooks containing all raw data
  • All computer generated data
  • Study correspondence
  • Statistical records
  • An original signed protocol, final report, an original signed report, the study correspondence and all raw data captured on durable media