Determination of an effective housekeeping gene for the quantification of mRNA for forensic applications |
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Authors: | Moreno Lilliana I Tate Courtney M Knott Erika L McDaniel Jade E Rogers Stephanie S Koons Barbara W Kavlick Mark F Craig Rhonda L Robertson James M |
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Institution: | Nuclear DNA Unit, FBI Laboratory Division, Quantico, VA 22135, USA. |
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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. |
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Keywords: | forensic science forensic biology serology messenger RNA real‐time polymerase chain reaction glyceraldehyde‐3‐phosphate dehydrogenase beta‐actin beta‐2 microglobulin cyclophilin A phosphoglycerate kinase 1 ribosomal protein large P0 |
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