Forensic Sci Sample Clauses

Forensic Sci. Int. 134:180-186 examined 1,547 cases from 1997-2000 containing 2,424 typed samples of which 163 (6.7%) contained a mixed profile with only 8 (0.3%) coming from more than two contributors minor Ratios of the various mixture components stay fairly constant between multiple loci enabling deduction of the profiles for the major and minor components Some mixture interpretation strategies involve using victim (or other reference) alleles to help isolate obligate alleles coming from the unknown portion of the mixture 95.1% (155/163) were 2-component mixtures xxxx://xxx.xxxx.xxxx.xxx/biotech/strbase/interlab/MIX05.htm Example Mixture Data (MIX05 Study-Profiler Plus) MIX05 Case #1; Profiler Plus green loci Single Source Sample (Victim) Evidence Mixture (Victim + Perpetrator) Amelogenin D8S1179 D21S11 D18S51 Obligate Alleles (not present in the victim reference) Y 12 28 16 True “Perpetrator” Profile X,Y 12,12 28,31.2 15,16 Victim = major Perpetrator = minor Mixtures: Issues and Challenges From X.X. Xxxxxx (2005) Forensic DNA Typing, 2nd Edition, p. 155 • The probability that a mixture will be detected improves with the use of more loci and genetic markers that have a high incidence of heterozygotes. • The detectability of multiple DNA sources in a single sample relates to the ratio of DNA present from each source, the specific combinations of genotypes, and the total amount of DNA amplified. • Some mixtures will not be as easily detectable as other mixtures. MIX05 Case #1; Identifiler green loci xxxx://xxx.xxxx.xxxx.xxx/biotech/strbase/interlab/MIX05.htm Mixture Mixture? Mixture? Mixture Mixture Mixtures: Issues and Challenges • Artifacts of PCR amplification such as stutter products and heterozygote peak imbalance complicate mixture interpretation • Thus, only a limited range of mixture component ratios can be solved routinely Our discussions have highlighted a significant need for continuing education and research into this area. ISFG Recommendations on Mixture Interpretation July 13, 2006 issue of Forensic Science International Two Parts to Mixture Interpretation • Deduction of alleles present in the evidence (compared to victim and suspect profiles) • Providing some kind of statistical answer regarding the weight of the evidence – An ISFG DNA Commission (Xxxxx Xxxx, Xxxxx Xxxx, Xxxxxxx Xxxxxxx, etc.) is evaluating the statistical approaches to mixture interpretation and has made recommendations Xxxx et al. (2006) DNA Commission of the International Society of F...
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Forensic Sci. Int. 164: 33-44. 6-FAM D10S1248 VIC D14S1434 XXX D22S1045 NC01 Loci NC Miniplexes NC01 NC02 NC03 D10S1248 D1S1677 D3S3053 D14S1434 D2S441 D6S474 D22S1045 26 total new loci NC04 NC05 NC06 NC10 D1GATA113 D1S1627 D3S4529 D3S3053 D2S1776 D8S1115 D9S2157 D6S474 D4S2408 D9S324 D10S1430 D20S482 NC07 NC08 NC09 D9S1112 D17S1301 D10S2327 D12ATA63 D18S8534 D11S4463 D14S1280 D20S1082 D17S974 Removed because they were problematic Why Build a Large Multiplex? For use with Reference Primer Concordance Checks/Null Alleles Samples Kinship Analysis Determine Mutation Rates Multiplex Complex Criminal Paternity Testing Immigration Testing Parentage Testing Missing Persons/ Mass Disasters The Multiplex Design Process – Xxxxxx, X.X., Xxxx, X.X., Xxxxxx, X.X., Xxxxx, M.C., Xxxx, X.X., Xxxxxxx, P.M. (2007) New autosomal and Y-chromosome STR loci: characterization and potential uses. Proceedings of the D4S2364 D20S482 Current Autoplex – 26 Loci 6 loci in VIC It didn’t work the first time… 7 loci in XXX X = 80 bp Y = 83 bp Amelogenin Xxxx-Xxxxxxxx and Xxxxxx (1997) Int J Legal Med 110(6): 312-315 6 loci in PET Sizes all <400 bp 7 loci in 6FAM The Design of the Multiplex • Goal: A single amplification 5-dye multiplex to combine the 26 new autosomal loci + Amelogenin in one reaction (27plex) • How can this be achieved? – Initial placement of all loci within 6FAM, VIC, NED, and PET dye channels (the size standard is in the LIZ channel) – Primer redesign for all but 7 of the original miniSTR loci – Trial and error of primer compatibility, as well as balancing for all working primers Some Loci Had to be Rearranged 100 removed 200 300 400 D6S474 D12ATA63 D22S1045 D10S1248 D1S1677 D4S2364 D9S1122 D2S1776 D10S1435 D3S3053 D9S2157 D3S4529 D2S441 D6S1017 D4S2408 D14S1434 D17S1301 D1GATA113 D18S853 D20S482 10 iterations later… D20S1082 D6S474 D12ATA63 D4S2364 D9S1122 D2S1776 D1S1627 D3S4529 D2S441 D22S1045 D10S1248 D1S1677 D11S4463 D10S1435 D3S3053 D5S2500 D17S974 D6S1017 D4S2408 D9S2157 D17S1301 D1GATA113 D18S853 D20S482 D14S1434 added later D17S974 D11S4463 D20S1082 D1S1627 Some redesign was necessary – certain complications occurred which will be further discussed… Amel D5S2500 D8S1115 These loci use the original miniSTR primers and thus have not been redesigned These loci use the original miniSTR primers and thus have not been redesigned Where do we begin? • Once the initial design has been laid out, primers were designed using Primer3 • Next, they were tested for multiplex-ability with A...
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Forensic Sci. Int. 148:1-14 TOTAL SAMPLES EXAMINED 1269 General Steps for Internal Validation • Review literature and learn the technique • Obtain equipment/reagents, if necessary • Determine necessary validation studies (there can be overlap and you only need to run a total of 50 samples) • Collect/obtain samples, if necessary • Perform validation studies maintaining all documentation • Summarize the studies and submit for approval to Technical Leader • Write-up the analytical procedure(s). Include quality assurance (controls, standards, critical reagents and equipment) and data interpretation, as applicable • Determine required training and design training module(s) • Design qualifying or competency test From Xxxxx Xxxxxxxx (FDLE), Validation Workshop (Aug 24-26, 2005 at NFSTC) xxxx://xxx.xxxx.xxxx.xxx/biotech/strbase/validation/validationworkshop.htm PowerPlex Y Developmental Validation Experiments Study Completed (17 studies done) Single Source (Concordance) Mixture Ratio (male:female) Description of Samples Tested (performed in 7 labs and Promega) 5 samples x 8 labs 6 labs x 2 M/F mixture series x 11 ratios (1:0,1:1,1:10,1:100,1:300,1:1000,0.5:300, 0.25:300,0.125:300, 0.0625:300, 0.03:300 ng M:F ) 6 labs x 2 M/M mixtures series x 11 ratios (1:0, 19:1, 9:1, 5:1, 2:1, 1:1, # Run 40 132 Revised SWGDAM Validation Guidelines (July 2004) xxxx://xxx.xxx.xxx/hq/lab/fsc/backissu/july2004/standards/2004_03_standards02.htm The document provides validation guidelines and definitions approved by SWGDAM July 10, 2003.
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