Authentication of cell lines is very important in biomedical research NIH has defined guidelines for authentication of any cell line to be used in grant eligible projects and many journals now require confirmation.
When should cell line authentication be conducted?
Authentication is required every time a new line is established or acquired.
Cells should be authenticated again before freezing, once every two months that the culture is actively growing, at any point that the performance of the line is inconsistent or yields unexpected results, and before publication.
Test cells whenever more than one cell line is in active use in the same laboratory to rule out cross-contamination.
Test cells when beginning any new series of experiments.
Our facility makes STR profiling of Human Cell Lines fast, easy and reliable. We offer 2 options for our users, Run Ready-PCR is prepared in your lab using either GenePrint® 10 System or PowerPlex® 16 HS System and sent to us, or Full Service- send us extracted DNA and we perform the PCR for you.
For both options, data is returned in an easy to understand format. This includes allele calls for each locus, screen shots of the raw data for comparison, and comparisons using the human STR profile database which includes data sets of 2455 cell lines from ATCC, DSMZ, JCRB and RIKEN.
Doing Good Science: Authenticating Cell Line Identity (article link)
- Extract DNA from cells using any commercial kit following manufacturer’s instructions. We recommend the Promega Wizard(R) Genomic DNA Purification Kit.
- After extraction, use a Nanodrop to calculate concentration and purity. The concentration must be at least 10ng/ul and purity should be in the range of 1.8-2.0 260/280nm. See this Technical Bulletin regarding assessment of Nucleic Acid purity.
Run-Ready Option (Rates)
- Prepare a PCR reaction following the protocol provided for the GenePrint® 10 System or PowerPlex® 16 HS System. We highly recommend including a positive control (2800M comes in each kit from Promega) and a negative control. The quality of purified DNA, small changes in buffers, ionic strength, choice of thermal cycler and thermal cycling conditions can affect PCR success. We suggest strict adherence to recommended procedures for amplification.
- Send 10ul of PCR product for samples and controls. Keep the remaining volume as back up.
Full Service Option using GenePrint® 10 System (Rates)
Send purified DNA with a concentration of at least 10ng/ul, purity should be in the range of 1.8-2.0, 260/280nm. If a Nanodrop is not available, that’s okay, we run your samples on our Nanodrop when they arrive. If sample purity or concentration is not in the correct range, we’ll contact you. See this Technical Bulletin regarding assessment of Nucleic Acid purity.