Determination of an Effective Housekeeping Gene for the Quantification of mRNA for Forensic Applications


  • This is publication number 10-21 of the Federal Bureau of Investigation (FBI). All samples were collected from consenting individuals according to FBI’s institutional standards. Names of commercial manufacturers are provided for identification purposes only, and inclusion does not imply endorsement of the manufacturer, its products, or services by the FBI. The views expressed are those of the authors and do not necessarily reflect the official policy or position of the FBI or the U.S. Government. This research was supported in part by the FBI’s Visiting Scientist Program, an educational opportunity offered by the Oak Ridge Institute for Science and Education (ORISE).

Additional information and reprints requests:
James M. Robertson, Ph.D.
Research Biologist
FBI Laboratory
2501 Investigation Parkway, Rm 4250
Quantico, VA 22135


Abstract:  The potential application of mRNA for the identification of biological fluids using molecular techniques has been a recent development in forensic serology. Constitutively expressed housekeeping genes can assess the amount of mRNA recovered from a sample, establish its suitability for downstream applications, and provide a reference point to corroborate the identity of the fluid. qPCR was utilized to compare the expression levels of housekeeping genes from forensic-like body fluid stains to establish the most appropriate assessment of human mRNA quantity prior to profiling. Although variability was observed between fluids and individuals, results indicated that beta-2 microglobulin exhibited the highest expression for all body fluids examined and across donors. A one-way analysis of variance was performed for housekeeping gene variability between donors (at the α, 0.05, significance level), and the results indicated significant differences for semen, vaginal secretions, and menstrual blood.