The JH-Nucleic Acid Technologies lab offers Short Tandem Repeat (STR) profiling for cell line authentication
Our services use STR profiles 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.
Authentication is recommended:
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when a new line is established or acquired.
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before freezing.
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Every two months that the culture is actively growing.
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if the performance of the line is not consistent or results are unexpected .
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before publication.
Cell Line Authentication Service Choices
The JH-Nucleic Acid Technologies Lab offers two levels of service support for STR profiles.
Standard Service
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
Standard Service plus Profile Search includes the Standard Service, plus profile comparison to public databases maintained by recognized repositories ATCC and DSMZ to check the authenticity of your cell line.
Cell line authentication analysis following ANSI/ATCC ASN-0002-2001 , the standardization of STR profiling using multiple kits.
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.
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.
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.
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.
Please inquire about DNA extraction from Non-Human sources
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 within 2 weeks of receipt.
Johns Hopkins Customers
Drop off samples in Blalock 1004 by 4PM on Monday for inclusion in that week’s processing schedule. If a sample needs to be repeated it will be included the next time the assay is run. Johns Hopkins customers should drop off samples in Blalock 1004 by 4PM on Monday for inclusion in that week’s processing schedule.
External Customers
Ship samples to the lab to be received by 11AM Tuesday for inclusion in that week’s processing schedule. Ship at ambient temperature or with ice packs. Ship samples with the customer information page of the request form to:
GRCF, Johns Hopkins University, 725 N. Wolfe Street, PCTB G20, Baltimore, MD 21205
Phone (443) 287-7948
Email the courier tracking number and request form to FAF@jhmi.edu.
STR Profile Analysis only
Standard Service:
GenePrint 10: $110/sample.
PowerPlex 18D: $140/sample.
Standard Service Plus Profile Search:
GenePrint 10: $155/sample.
PowerPlex 18D: $185/sample.
NOTE:
For this service DNA samples can be submitted. If DNA isolation is required (cell pellets samples): $25 per sample is the fee for isolation.
EZ-PCR Mycoplasma test and Str profile analysis
GenePrint 10: $170/sample (Standard Service).
GenePrint 10: $215/sample (Standard Service plus Profile Search).
PowerPlex 18D: $200/sample (Standard Service).
PowerPlex 18D: $245/sample (Standard Service plus Profile Search).
NOTE: DNA isolation fee is included in in the mycoplasma test.
Mycodtect mycoplasma test and str profile service
GenePrint 10: $180/sample (Standard Service).
GenePrint 10: $225/sample (Standard Service plus Profile Search).
PowerPlex 18D: $210/sample (Standard Service).
PowerPlex 18D: $255/sample (Standard Service plus Profile Search).
NOTE: DNA isolation fee is included in in the mycoplasma test.
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