법과학 증거물들에 대한 PreCRTM 복구 효소의 응용
Application of PreCRTM Repair Enzyme to Forensic Evidences
0
조회
0
다운로드
0
인용
0
참고문헌
키워드
초록
DNA가 손상된 법과학 시료에서 성공적인 STR 분석을 하기 위해 PreCRTM 복구 효소를 적용하였다. 본 실험을 위해 총 122개 시료를 선별하였으며 복구 반응 후, Identifiler® plus 키트를 이용하여 STR 좌위 분석을 수행하였다. 또한 Quantifiler® 키트를 이용하여 복구반응 전, 후의 DNA를 정량하여 비교하였다. 그 결과, 89개 시료에서 STR allele의 복구 반응이 확인되었다. 복구반응 후 소실되었던 STR 좌위의 값이 증폭되었고, STR peak의 높이가 증가하였다. 그러나 몇 개의 시료에서는 비특이적 피크가 생기는 양상을 나타냈다. 이 중 42개 시료를 대상으로 DNA 정량을 수행해 본 결과, 18개 시료에서 복구 반응 후 DNA양이 증가하였다. 따라서 PreCRTM 복구효소는 다양한 환경에 노출된 법과학 시료에 효율적으로 적용할 수 있지만, 결과가 산발적이거나 오히려 저해되는 경우도 발생하기 때문에 복구반응 후 결과 분석에 주의를 기울여야 한다.
Abstract
PreCRTM repair enzyme was applied to a variety of damaged forensic samples to obtain a successful STR result. A total of 122 samples were selected for the analysis. We analyzed STR allele recovery with repair reaction and Identifiler ® plus amplification kit. Also, DNA quantification was performed with the Quantifiler ® kit. As results, 89 samples repaired with the PreCRTM. Some lost alleles restored and STR peak heights increased upon repair with this kit. In contrast, non-specific peaks were observed in some other samples. For 18 samples among 42 samples, DNA concentration was increased after the recovery. In conclusion, PreCRTM repair enzyme will be efficiently applied to forensic samples which were damaged by various environments. However, the results were sporadic and sometimes deleterious. Therefore, the results of STR typing after the enzyme treatment must be carefully analyzed.
참고문헌 (12)
- DH Warshauer, "STRait Razor: a length-based forensic STR allele-calling tool for use with second generation sequencing data", Forensic Sci Int Genet, 7(4), pp.409~417 (2013)
- JM Butler, "The development of reduced size STR amplicons as tools for analysis of degraded DNA", J Forensic Sci, 48(5), pp.1054~1064 (2003)
- R Alaeddini, "Forensic implications of PCR inhibition—A review", Forensic Sci Int Genet, 6(3), pp.297~305 (2011)
- C Thomas Evans Jr, "NEB Expressions-DNA damage", Ipswich: New England Biolabs, 2, pp.1~3 (2007)
- S Paabo, "Genetic analyses from ancient DNA", Annu Rev Genet, 38, pp.645~679 (2004)
- T Lindahl, "Rate of depurination of native deoxyribonucleic acid", Biochemistry, 11(19), pp.3610~3618 (2002)
- VI Bruskov, "Heat-induced formation of reactive oxygen species and 8-oxoguanine, a biomarker of damage to DNA", Nucleic Acids Res, 30(6), pp.1354~1363 (2002)
- P Gil, "An investigation of the rigor of interpretation rules for STRs derived from less than 100 pg of DNA", Forensic Sci Int, 112, pp.17~40 (2000)
- JM Butler, "Forensic DNA typing. Biology, technology and genetics of STR markers" (2005), Elsevier Academic Press
- JM Robertson, "Assessing PreCR™ repair enzymes for restoration of STR profiles from artificially degraded DNA for human identification", Forensic Sci Int Genet, 12, pp.168~180 (2014)
- AA Westen, "Degraded DNA sample analysis using DNA repair enzymes, mini-STRs and (tri-allelic) SNPs", Forensic Sci Int Genet Supplement, 2, pp.505~507 (2009)
- TM Diegoli, "An optimized protocol for forensic application of the PreCRTM Repair Mix to multiplex STR amplification of UV-damaged DNA", Forensic Sci Int Genet, 6(4), pp.498~503 (2012)