Molecular Biology Core Facility 

Human Identity Testing/STR Profiling

 

The MBCF performs the service of Human Identity testing for the purposes of zygosity testing and cell line authentication (CLA). We utilize the STR DNA Profiling Globalfiler PCR Amplification Kit from Thermo Fisher.

The MBCF provides STR profiling for the purposes of CLA for customers that wish to confirm the identity of a cell line. Identity can be ‘authenticated’ by comparison to a known cell line DNA profile or by comparing allelic information and matching/confirming identity with published information, commercial reference information, or a reference sample from the donor. The process of donor origin authentication is a multifaceted process with many elements of data analysis to reach a final conclusion. In an effort to aid users in implementation of the most current accepted methods and best scientific practices, we’ve assembled a list of very informative links below with very detailed information about utilizing your DNA ‘fingerprint’ once you’ve received your report from us. Alternatively, you can request that we perform the authentication analysis and provide you with a concise summary report of your tested sample.

Two STR Testing Services are available-

  1. Standard Testing- Once your samples have been processed by the MBCF, you will receive a STR profile table and electropherogram (graphic) profile for each of your samples.
  2. Standard Testing Plus Match Analysis- the Standard Testing and a Summary report of your sample. The Summary report includes Match Analysis of your sample profile and database search results of cell line repositories ATCC, DSMZ, JCRB, COG and NCBI BioSample. For Match Analysis, verifiable reference profile data must be obtainable or provided, or a reference sample can be provided for testing. A % match score is provided for authentication.
Please refer to our "Online Sample Submission" page for step-by-step instructions to enter your samples for testing.

    Cell Line Authentication Testing Guidelines/Recommendations
    These are published guidelines provided by the ATCC Standards Development Organization Workgroup (ASN-0002). 

    Authentication testing at key intervals of a project should be performed when you-

    1. Establish or acquire a new cell line. 
    • At an early passage (within first week of culture).
    • From donor tissue if possible.
    • Testing not necessary if acquired from a trusted sources such as ATCC.​
     2. Start a new series of experiments.
    ​ 3. Are routinely passaging cell lines.
    • Every 10 passages.
     4. See inconsistent cell behavior or unexpected results.
     5. Prepare to publish or submit grant application.
     6. Freeze cell stocks.

    Links for STR Cell Line Authentication Resources and Documentation

    Implementing Rigor and Transparency in NIH & AHRQ Research Grant Applications.
    http://grants.nih.gov/grants/guide/notice-files/NOT-OD-16-011.html

    NIH Notice regarding CLA.
    http://grants.nih.gov/grants/guide/notice-files/NOT-OD-08-017.html

    ICLAC Guide to CLA.
    http://standards.atcc.org/kwspub/home/the_international_cell_line_authentication_committee-iclac_/Authentication_SOP.pdf

    ICLAC Advice to Scientists.
    http://standards.atcc.org/kwspub/home/the_international_cell_line_authentication_committee-iclac_/Advice_to_Scientists.pdf

    ATCC ICLAC.
    http://standards.atcc.org/kwspub/home/the_international_cell_line_authentication_committee-iclac_/

    ATCC Cell Line Authentication Test Recommendations.
    http://www.atcc.org/~/media/PDFs/Technical%20Bulletins/tb08.ashx

    Guidelines for the Use of Cell Lines in Biomedical Research.
    http://www.nature.com/bjc/journal/v111/n6/full/bjc2014166a.html

    Match criteria for human cell line authentication: Where do we draw the line?
    Int J Cancer. 2013 Jun 1;132(11):2510-9. doi: 10.1002/ijc.27931. Epub 2012 Nov 28.

    Recommendation of short tandem repeat profiling for authenticating human cell lines, stem cells, and tissues.
    In Vitro Cell Dev Biol Anim. 2010 Oct;46(9):727-32. doi: 10.1007/s11626-010-9333-z. Epub 2010 Jul 8.

    Short tandem repeat profiling: part of an overall strategy for reducing the frequency of cell misidentification.
    In Vitro Cell Dev Biol Anim. 2010 Dec;46(10):811-9. doi: 10.1007/s11626-010-9352-9. Epub 2010 Oct 7.

    ATCC resources page- scroll down to CLA links.
    http://www.atcc.org/Documents/Learning_Center/Resources_for_Cell_Biology.aspx

    BioSample- List of published cell line data. 
    http://www.ncbi.nlm.nih.gov/biosample/

    Promega CLA reference List.
    http://www.promega.com/products/pm/cell-authentication/references/

    Mycoplasma Testing for Biologics from WuXi AppTec.
    www.wuxiapptec.com/pdfs/WA-MycoplasmaGuide.pdf

    Location: 
    Barbara Davis Center
    University of Colorado, Anschutz Medical Campus
    1775 Aurora Court, 4th floor, Room 4207
    Aurora, CO 80045 
    Phone: 303-724-6825

    Molecular Biology contact personnel:
    Mark Farrell​, Core Manager


     

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