TUĞBA ÜNSAL SAPAN, GÖNÜL FİLOĞLU TÜFEK, Havva ALTUNÇUL, Kadir DASTAN, ÖZLEM BÜLBÜL

Çeşitli yüzeyler üzerindeki kan lekelerinde 6 mini str lokusunun (D1S1677, D2S441, D4S2364, D10S1248, D14S1434, D22S1045) profillendirilmesi

Klasik STR lokusları ile olay yerinden gelen aşırı derecede bozulmuş biyolojik örneklerde ya tipleme sorunları yaşanmakta ya da sonuç alınamamaktadır. Bunu ortadan kaldırmaya yönelik, National Institute of Standards and Technology (NIST) tarafından 26 yeni mini STR lokusu geliştirilmiştir. Bu lokusların PCR ürünleri 150 bç'den küçük olup özellikle de 6 lokusun (D1S1677, D2S441, D4S2364, D10S1248, D14S1434, D22S1045) PCR ürünleri 125 bç'den küçüktür. Bu çalışmada olay yerinde bulunabilecek çeşitli yüzeylere (kot, penye, döşeme, havlu, bıçak kabzası, laminant parke, kapı kilit demiri ve kalebodur arası dolguderz) kan damlatılarak 1 hafta, 1 ay, 3 ay ve 6 ay bekletildi. Bu lokuslar için daha önceden optimizasyon ve validasyon çalışmasında belirlenen PCR koşulları uygulandı [8]. 1 hafta, 1 ay, 3 ay ve 6 aylık kan lekelerinde 6 miniSTR lokusu tiplendirildi. Yapılan hassasiyet çalışması sonucunda 6 mini STR lokusuna ait analizlerde kullanılabilecek en az DNA miktarının 0.1 ng/?l olduğu belirlendi

Profiling of 6 mini STR loci (D1S1677,D2S441,D4S2364, D10S1248, D14S1434, D22S1045) on blood spots waited on various surfaces

Conventional STR loci are insufficient for successful analysis of DNA specimens from mass disasters or forensic evidence. In order to solve this problem, The National Institute of Standards and Technology (NIST) developed 26 new STR loci. The PCR products of these loci are less than 150 bp in size. Especially the PCR products of 6 loci (D1S1677, D2S441, D4S2364, D10S1248, D14S1434, D22S1045) are less than 125 bp in size. In this study, blood was dripped on various surfaces that may be found in crime scenes (jean, t-shirt, couch upholstery, towel, knife hilt, laminate parquet, door lock iron and grouting between tiles) and they were kept waiting for 1 week, 1 month, 3 months and 6 months. For the mentioned loci, the PCR conditions previously determined in the optimization and validation process were applied [8]. At the end of this study, it was determined that the minimum DNA quantity of 6 mini SRT loci can be 0.1 ng/μl for identification of blood spots

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1. Butler JM, Shen Y, McCord BR. The Development of Reduced Size STR Amplicons as Tools for Analysis of Degraded DNA. J Forensic Sci Int. 2003;48(5):1054-64.
2. Butler JM. Short tandem repeat typing technologies used in human identity testing. Bio Techniques. 2007;43(4):2-4.
3. Coble MD, Butler JM. Characterization of New MiniSTR Loci to Aid Analysis of Degraded DNA. J Forensic Sci. 2005;50:43-53.
4. Hill CR, Coble MD, Butler JM. Development of 27 New MiniSTR Loci for Improved Analysis of Degraded DNA Samples. American Academy of Forensic Sciences, Seattle, WA. 2006; Poster B105.
5. Hill CR, Coble MD, Butler JM. Characterization of 26 New MiniSTR Loci. Promega Meeting, Nashville, TN. 2006; Poster 44.
6. Carboni I, Lozzi S, Nutini AL, Torricelli F, Ricci U. Improving complex kinship analyses with additional STR loci. Elektrophoresis. 2014;35(21-22):3145-51.
7. NIST mini STR homepage
8. Unsal T, Filoglu G, Sipahi E, Rayimoglu G, Altuncul H. New MiniSTR Loci D10S1248, D14S1434, D22S1045, D4S2364, D2S441, D1S1677 Validation And Optimization On Blood Samples. Forensic Science International: Genetics Supplement Series. 2011;3:e473-e474.
9. Weber JL, May PE. Abundant class of human DNA polymorphisms which can be typed using the polymerase chain reaction. Am J Hum Genet. 1989;4483):388-96.
10. Edwards A, Hammond HA, Jin L, Caskey T, Chakraborty R. Genetic variation at five trimeric and tetrameric tandem repeats. Genomics. 1992;12(2):241-53.
11. Hill CR, Kline MC, Coble MD, Butler JM. Characterization of 26 MiniSTR Loci for Improved Analysis of Degraded DNA Samples. J Forensic Sci. 2008;53(1):73-80.
12. Constantinescu CM, Barbarii LE, Ianc CB, Constantinescu A, Neagu E, Iancu D, Girbea G. Challenging DNA samples solved with MiniSTR analysis. Rom J Leg Med. 2012;20:51-6.
13. National DNA Index System (NDIS) Operational Procedures -FBI https://www.fbi.gov/about-us/lab/biometric-analysis/codis/pla access date 21.06.2016
14. McCord B, Opel K, Funes M, Zoppis S, and Jantz LM. An Investigation of the Effect of DNA Degradation and Inhibition on PCR Amplification of Single Source and Mixed Forensic Samples. U.S. Department of Justice Final Report. 2011;2006-DN-BX-K006.