Cell Line Identity Validation – STR Profiling for HUMAN Cell Line Authentication
Delivery of samples      Ordering         Pricing                Questions:Laura Kasch [410-614-3830]
Sample preparation      Turnaround   What you will receive

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

DNA Services provides cell line authentication to the Hopkins community and any academic or research facility.

General recommendations are to authenticate:

  • when a new line is established or acquired
  • 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

DNA Services offers two levels of human cell line authentication service, Standard Service or Standard Service plus Profile Search. Standard Service includes a signed cell line authentication report for each line submitted and is for scientists who prefer to do their own profile comparison analysis. The report contains a STR profile table, an electropherogram profile, and a table with the raw data for each allele (size in base pairs, peak height, peak area). The results can be used for comparison to reference databases to check for cross contamination, mix-ups and percent match between lines or to establish the identity of a new cell line. The certificate is suitable for proof of cell line authentication required by journals and grant funding bodies. Standard Service plus Profile Search includes the above, plus profile comparison to public databases maintained by recognized repositories ATCC and DSMZ to check the authenticity of your cell line.

GRCF DNA Services offers cell line authentication analysis following ANSI/ATCC ASN-0002-2011, Authentication of Human Cell Lines: Standardization of STR Profiling guidelines using multiple kits. The profiling kits are only suitable for cell lines of HUMAN origin. A Beta-globin mouse marker option is offered to test for the presence of mouse DNA.

  • GenePrint 10 System (Promega). Markers included in the kit are amelogenin, CSF1PO, D13S317, D16S539, D21S11, D5S818, D7S820, THO1, TPOX, and vWA. This kit contains all the markers listed in ATCC’s database plus D21S11.
  • PowerPlex 18D (Promega): Markers included in the kit are amelogenin, CSF1PO, D13S317, D16S539, D21S11, D5S818, D7S820, THO1, TPOX, vWA, D18S51, D3S1358, FGA, D8S1179, Penta E, Penta D, D2S1338 and D19S433. This kit contains all the markers listed in ATCC’s database plus 9 additional markers not listed. This kit is most similar to the Identifiler (Applied Biosystems) kit that is no longer available for research purposes.
Ordering:

Johns Hopkins customers please place your order through our secure online ordering system, Genesifter. Genesifter is now behind the JHMI firewall. This means it is ONLY accessible within the Hopkins environment. If you are off campus you will need to access the server via VPN, just like your email and many other JHU secure services. For the order form see FAF Order Forms: FAF Cell Line Authentication (click on the image).

External non-Johns Hopkins customers please use this downloadable Excel order form for placing requests. Our online ordering system, Genesifter, is no longer available to external customers due to security risks. We hope to have a solution in the future. If you have a Genesifter account with us and need access to your old data files please contact Laura Kasch at lkasch@jhmi.edu. Sorry for the inconvenience.

Sample preparation for human Cell Line Authentication
The GRCF (FAF) accepts purified DNA (please provide >10µl at >10ng/µl of DNA), cell pellets (dried or frozen, between 100,000 and 5 million, preferred, pellet should be visible) or frozen cell aliquots (Cryopreservation tube) for analysis. A visible cell pellet is required for combined authentication and mycoplasma testing. A DNA isolation charge applies when submitting cell pellets or frozen cell aliquots for authentication service. We can also extract DNA from other sources – please inquire.

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Delivery of samples

JH Customers:

Drop samples off in Blalock 1004 with the GeneSifter order number, Monday – Friday from 9 AM – 4:00 PM. All sample types may be delivered at ambient temperature.

External Customers:

All sample types may be shipped at ambient temperature or with ice packs, dry ice is not required. Ship samples with the GeneSifter order number to:

Laura Kasch
Fragment Analysis Facility
Johns Hopkins University
2760 Lighthouse Point East, Suite 201
Baltimore, MD 21224
Phone (443) 287-7948

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).

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, a graphic profile (electropherogram), and a table with the raw data for the alleles with base pair size, peak height, and peak area. The signed report provides a STR profile for the cell line submitted, it does not automatically authenticate the cell line. Authentication is determined by comparison to the STR profile of the original donor tissue or repository cell stock. The STR profile is used to verify the cell line is human, evaluate profile consistency between provisionally related cell isolates, detect cross-contamination with another human cell line (intra-species contamination), and can be used to compare to profile databases, which we strongly recommend. Additional information is provided with links to profile databases and information about interpreting your cell line results. The report is suitable for proof of cell line authentication that may be required by journals and grant funding bodies, though you will need to check the profile of your line against published profiles. If preferred, the GRCF provides databases searches with our Plus Profile Search service. 

Standard Service Plus Profile Search: You will receive a signed cell line authentication report that includes a STR profile table of your cell line, an graphic profile (electropherogram), and a table with the raw data for the alleles with base pair size, peak height, and peak area, plus a database search summary of repositories ATCC and DSMZ. The report is suitable for proof of cell line authentication that may be required by journals and grant funding bodies.

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. If a samples fails processing it will be repeated the following week.

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2018 Pricing

DNA isolation (required for submission of cell pellets): $20 per sample

STR Profiling Analysis:

Standard Service:

  • GenePrint 10: $100/sample
  • PowerPlex 18D: $140/sample

Standard Service Plus Profile Search:

  • GenePrint 10: $142/sample
  • PowerPlex 18D: $182/sample

Mycoplasma testing AND human cell line authentication (STR profiling) from the same CELL PELLET (a cell pellet is needed for combined service, extracted DNA is not suitable):

  • GenePrint 10: $145/sample (Standard Service)
  • GenePrint 10: $180/sample (Standard Service plus Profile Search)
  • PowerPlex 18D: $179/sample (Standard Service)
  • PowerPlex 18D: $214/sample (Standard Service plus Profile Search)
Payment options:

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

International customers using a credit card will have an additional $25 processing fee added.