Animal Cell Line Authentication by DNA Barcoding

DNA barcoding will determine the species of a cell line. It involves amplifying and sequencing mitochondrial DNA. Mitochondrial DNA shows 5–10-fold more differences between species than nuclear DNA. The genetic analysis of mitochondrial genome sequences is simpler than the analysis of nuclear DNA because only a single PCR and sequencing reaction are normally required; mitochondrial DNA shows only very low within-species variation.

How to submit samples

Please complete the two forms below and send to us  using the address below prior to arranging shipment of samples:

• Sample Submission Form
• Biohazard Risk Assessment

Once the forms have been received, reviewed and accepted by ECACC, you will be issued with a reference number that will need to be quoted on your samples and in all future correspondence. You will also need to provide a purchase order number.

If sending DNA:

• DNA should be sent in an o-ring sealed centrifuge tube.  If an o-ring sealed tube is not available, then seal the tube lid with Parafilm to avoid leakage or loss of liquid by evaporation
• DNA should be at a concentration of 10ng/µl and a minimum volume of 50µl
• DNA should be sent on ice packs

If sending growing cultures:

• growing cultures should be sent in flasks topped up with medium. The flask cap should be of the non-vented variety and wrapped e.g. in Parafilm, to prevent leakage
• growing cultures for STR profiling (human cell lines only) can be sent as a viable culture with at least 2 x 10⁶ cells per container

If sending frozen cultures:

• a handling fee will be charged to cover costs of establishing the cell lines in culture where required (not relevant to samples for STR profiling where the DNA can be extracted from frozen ampoules containing at least 2 x 10⁶ cells/amp)
• details of the cell line growth media and culture conditions must be supplied. Where specialised media or supplements are used you may be asked to supply sufficient quantities to establish the lines in culture

If sending PBS washed cell pellets:

• phosphate buffered saline (PBS) washed pellets submitted for STR profiling must contain at least 2 x 10⁶ cells

Flasks and ampoules should be adequately packed and transported according to UN602 regulations. If you use a courier service to ship your samples, please contact the company involved for advice on their own approved packaging. ECACC will be happy to return any packaging to you for re-use.

Parcels should be addressed to:

QC Testing Department Culture Collections Public Health England Porton Down Salisbury SP4 0JG UK

Notification on dispatch of samples to ECACC is advised, please email us on culturecollections@phe.gov.uk