Determination of an effective housekeeping gene for the quantification of mRNA for forensic applications.
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.
Nuclear DNA Unit, FBI Laboratory Division, Quantico, VA 22135, USA.
Lilliana I
LI
Moreno
Courtney M
CM
Tate
Erika L
EL
Knott
Jade E
JE
McDaniel
Stephanie S
SS
Rogers
Barbara W
BW
Koons
Mark F
MF
Kavlick
Rhonda L
RL
Craig
James M
JM
Robertson
D000199:Actins / Q000235:genetics; D000328:Adult; D000704:Analysis of Variance; D001774:Blood Chemical Analysis; D002582:Cervix Mucus / Q000737:chemistry; D021984:Cyclophilin A / Q000235:genetics; D005260:Female; D053803:Forensic Genetics; D015870:Gene Expression; D020869:Gene Expression Profiling; D005986:Glyceraldehyde 3-Phosphate / Q000235:genetics; D006801:Humans; D008297:Male; D008598:Menstruation; D008875:Middle Aged; D010735:Phosphoglycerate Kinase / Q000235:genetics; D012333:RNA, Messenger / Q000378:metabolism*; D060888:Real-Time Polymerase Chain Reaction; D012269:Ribosomal Proteins / Q000235:genetics; D012463:Saliva / Q000737:chemistry; D012661:Semen / Q000737:chemistry; D001613:beta 2-Microglobulin / Q000235:genetics
D000199:Actins; D012333:RNA, Messenger; D012269:Ribosomal Proteins; D001613:beta 2-Microglobulin; C054675:ribosomal protein P0; D005986:Glyceraldehyde 3-Phosphate; D010735:Phosphoglycerate Kinase; D021984:Cyclophilin A