Cell Line Authentication

STR Profiling for Human Cell Line Authentication

News

Due to a US District Court ruling, Applied Biosystems is no longer able to sell their Identifiler kit for cell line authentication. The GRCF still offers authentication using Promega's StemElite kit.

STR Profiling

In light of the NIH's recommendation that all cell lines be authenticated before publication, the Fragment Analysis Facility offers a short tandem repeat (STR) profiling service to customers who wish to confirm the identity of a line by comparison to a known profile or to establish an identity profile for a new cell line generated locally.

The FAF provides services to the Hopkins community, and any academic or research facility.

Cell lines came become altered by contamination from another cell line or, due to their instability, from spontaneous genetic change. General recommendations are to authenticate:

when a new line is established or acquired to determine an identity for the cell line.
before freezing.
every two months that the culture is actively growing.
if the performance of the line is not consistent or results are unexpected.
before publication.

The FAF is currently offering two levels of service, Standard Service or Standard Service plus Profile Search. Standard Service includes a signed cell line authentication report for each line submitted, which is suitable for proof of cell line authentication that may be required by journals and grant funding bodies, an STR profile table with data suitable for comparison to reference databases, an electropherogram profile, and a table with the raw data for the alleles (size in base pairs, peak height, peak area). Standard Service is suitable for scientists who prefer to do their own profile comparison analysis. The results can be used to check for cross contamination, mix-ups and percentage of identity between lines or to establish the identity of a new cell line. Standard Service plus Profile Search includes the above, plus cross referencing against repositories ATCC, DSMZ, and JCRB databases to check the authenticity of your cell line.

The FAF is currently offering analysis using the StemElite ID System (Promega). The kit is only suitable for cell lines of human origin. Markers included in the kit are amelogenin, CSF1PO, D13S317, D16S539, D21S11, D5S818, D7S820, THO1, TPOX, and vWA. The StemElite ID System also includes a mouse specific marker for detection of cross contamination between mouse and human cell lines. See the below table for STR markers included in the kit.

Ordering

Please place your order through our new secure online ordering system. Click on "Create New Account" to create a login.

For assistance filling out the online request please call 443-287-7948 or view our CLA Ordering Instructions.

Delivery of samples

JH Customers: Drop samples off in Blalock 1004 with the GeneSifter order number, Monday - Friday from 9 AM - 4:00 PM.

External Customers:

Send the samples with the GeneSifter order number to:

Email the courier tracking number and GeneSifter order number to FAF@jhmi.edu. Samples must be received by 11AM Tuesday for inclusion in our weekly processing schedule (see Turnaround Time).

Sample preparation

The FAF accepts purified DNA (please provide a minimum of 100ng of DNA), cell pellets (dried or frozen, between 100,000 and 5 million, preferred) or frozen cell aliquots (Cryopreservation tube) for analysis. A DNA isolation charge applies when submitting cell pellets or frozen cell aliquots. We can also extract DNA from other sources - please inquire.

What you will receive

Standard Service: You will receive a signed cell line authentication report that includes a STR profile table of your cell line with data suitable for comparison to reference databases, an electropherogram (graphic profile), and a table with the raw data for the alleles (size in base pairs, peak height, peak area). The report is suitable for proof of cell line authentication that may be required by journals and grant funding bodies.

Standard Service Plus Profile Search: You will receive a signed cell line authentication report that includes a STR profile table of your cell line with data suitable for comparison to reference databases, an electropherogram (graphic profile), and a table with the raw data for the alleles (size in base pairs, peak height, peak area) plus a database search of repositories ATCC, DSMZ, and JCRB for a known cell line reference profile for comparison.

Turnaround Time

Samples submitted for STR profiling are run once a week on Tuesday, with results emailed usually on Monday of the following week. Johns Hopkins customers should drop off samples in Blalock 1004 by 4PM on Monday or hand deliver to the FAF laboratory in Canton by 11AM Tuesday for inclusion in that week's processing schedule. External customers should ship samples to the lab to be received by 11AM Tuesday for inclusion in that week's processing schedule. Samples received after 11AM Tuesday through Friday of any given week will be held and processed the following week.

Pricing*

DNA isolation (required for submission of cell pellets): $25 per sample

STR Analysis:

Standard Service:

StemElite: $80/sample

Standard Service Plus Profile Search:

StemElite: $120/sample

Mycoplasma AND cell line authentication (STR profiling) from the same cell pellet:

StemElite: $126/sample (Standard Service)
StemElite: $166/sample (Standard Service plus Profile Search)

Payment:

John Hopkins investigators: IO number or credit card (VISA or MasterCard)
External U.S. customers: P.O. or credit card (VISA or MasterCard)
External foreign customers: P.O., check or credit card (VISA or MasterCard)

*Listed pricing is for Johns Hopkins investigators; all other customers will have a 16% administrative fee added. International customers using a credit card will have an additional $25 processing fee added in addition to the administrative fee.

FAF services may only be used for research purposes. Any data or samples generated by the facility may not be used for human diagnostic or therapeutic use.