Methods, panels of identification markers, and kits for identifying forensic samples
View Patent ↗Methods for identifying forensic samples using panels of markers and gene expression profiling, including without limitation, mRNA profiling, miRNA profiling, or both, are disclosed. Panels of markers for identifying certain tissue samples and certain body fluid samples are also disclosed. Kits for expediting performance of certain of the disclosed methods are provided.
1. A set of primers specific for a panel of marker genes, wherein the set consists of primer pairs to specifically amplify each of the genes in the panel of marker genes, wherein the panel of marker genes includes the marker genes PRB4, MCSP, ANK1, ESR1, and MMP11 and wherein the set of primers consists of 21 or fewer primer pairs.
2. The set of primers specific for a panel of marker genes of claim 1 , optionally including one or more primer pairs to specifically amplify one or more marker genes selected from PRB1, PRB3, HTN3, SEMG1, SEMG2, TGM4, PRM1, PRM2, and MMP7.
3. The set of primers specific for a panel of marker genes of claim 1 , wherein the panel of marker genes includes at least one or more additional marker genes for a body fluid sample minimally including saliva, semen, blood, vaginal secretions, and menstrual blood.
4. The set of primers specific for a panel of marker genes of claim 3 , wherein the panel of marker genes includes one or more additional marker genes for a body fluid sample selected from saliva, semen, saliva containing mucus, vaginal secretions, blood, and menstrual blood.
5. A set of primers specific for a panel of marker genes, wherein the set consists of primer pairs to specifically amplify each of the genes in the panel of marker genes, wherein the panel of marker genes consists of the marker genes PRB4, MCSP, ANK1, ESR1, and MMP11 and optionally includes one or more primer pairs to specifically amplify one or more marker genes selected from PRB1, PRB3, HTN3, SEMG1, SEMG2, TGM4, PRM1, PRM2, and MMP7.
6. A method for identifying a forensic sample containing nucleic acid comprising:
combining a set of primer pairs to specifically amplify each of the genes in a panel of marker genes, wherein the panel of marker genes includes the marker genes PRB4, MCSP, ANK1, ESR1, and MMP11 with at least some of the nucleic acid from the sample and a polymerase;
amplifying the nucleic acid from the sample with the set of primer pairs, wherein the set of primers consists of 21 or fewer primer pairs;
generating an expression profile for the panel of marker genes; and
identifying the forensic sample.
7. The method of claim 6 , optionally including one or more primer pairs to specifically amplify one or more marker genes selected from PRB1, PRB3, HTN3, SEMG1, SEMG2, TGM4, PRM1, PRM2, and MMP7.
8. The method of claim 6 , wherein the amplifying comprises a polymerase chain reaction.
9. The method of claim 6 , wherein the generating comprises using a reporter probe, a substrate, a nucleic acid dye, a microfluidics device, or combinations thereof.
10. The method of claim 6 , wherein the polymerase comprises a DNA-dependent DNA polymerase or an RNA-dependent DNA polymerase and a DNA-dependent DNA polymerase.
11. The method of claim 6 , wherein the amplifying comprises a first amplification reaction comprising a multiplicity of different gene-specific primer pairs and a second amplification reaction comprising a single-plex amplification reaction.
12. The method of claim 11 , wherein the second amplification reaction comprises a multiplicity of different single-plex amplification reactions performed in parallel.
13. The method of claim 12 , wherein the multiplicity of parallel single-plex amplification reactions occur in a microfluidics device.
14. The method of any one of claims 6 - 13 , wherein the forensic sample is a body fluid sample.
15. The method of claim 14 , wherein the body fluid sample is selected from blood, semen, saliva, vaginal secretions, and menstrual blood.
16. A kit comprising of a set of primers specific for a panel of marker genes, wherein the set of primers consists of primer pairs to specifically amplify each of the genes in the panel of marker genes, wherein the panel of marker genes includes the marker genes PRB4, MCSP, ANK1, ESR1, and MMP11, wherein the set of primers consists of 21 or fewer primer pairs.
17. The kit of claim 16 , optionally including one or more primer pairs to specifically amplify one or more marker genes selected from PRB1, PRB3, HTN3, SEMG1, SEMG2, TGM4, PRM1, PRM2, and MMP7.
18. The kit of claim 16 , further comprising one or more of a polymerase; nucleotide triphosphates (NTPs), wherein the NTPs can be ribonucleotide triphosphates (rNTPs) and/or deoxyribonucleotide triphosphates (dNTPs); a reporter probe; a nucleic acid dye; a miRNA linker probe; an internal reference dye; a control sequence or internal standard; or combinations thereof.
19. The kit of claim 16 , further including a microarray and/or a microfluidics device.