FoLT-PCR에 의한 유전자형 (HumTH01, HumTPOX, HumCSF1PO & Amelogenin) 분석

Quadruplex Genotype Analysis at HumTH01, HumTPOX, HumCSF1PO and Amelogenin Loci by FoLT-PCR

  • 이양한 (국립과학수사연구소 유전자분석실) ;
  • 임시근 (국립과학수사연구소 유전자분석실) ;
  • 강필원 (국립과학수사연구소 유전자분석실) ;
  • 최동호 (국립과학수사연구소 유전자분석실) ;
  • 윤송노 (국립과학수사연구소 유전자분석실) ;
  • 한면수 (국립과학수사연구소 유전자분석실)
  • Lee, Yang-Han (DNA Analysis Section, National Institute of Scientific Investigation) ;
  • Lim, Si-Keun (DNA Analysis Section, National Institute of Scientific Investigation) ;
  • Kang, Pil-Won (DNA Analysis Section, National Institute of Scientific Investigation) ;
  • Choi, Dong-Ho (DNA Analysis Section, National Institute of Scientific Investigation) ;
  • Yoon, Song-Ro (DNA Analysis Section, National Institute of Scientific Investigation) ;
  • Han, Myun-Soo (DNA Analysis Section, National Institute of Scientific Investigation)
  • 투고 : 1999.01.21
  • 발행 : 1999.06.25

초록

법적시료(Forensic Evidences)로 사용되는 혈흔, 정액반, 타액반 및 모발을 DNA의 분리과정 없이 FoLT (Formamide Low Temperature) PCR법으로 DNA의 특정부위를 분석하고자 하였다. FoLT PCR법으로 3종류의 STR(Short Tandem Repeat) 좌위와 성별을 확인하는 Amelogenin gene을 PCR법으로 동시에 분석하기 위하여 formamide농도와 annealing온도를 검토한 바 최적의 formamide농도는 8%(v/v)이었고 annealing온도는 $48^{\circ}C$이었다. 그리고 1% Triton X-100을 이용하여 시료를 세척한 경우에 증폭 효율이 증가하였다. 따라서 FoLT-PCR법을 이용한 경우 DNA를 분리하기 위한 전처리없이 소량의 시료를 기존의 PCR법보다 빠른 시간내에 한번의 PCR 및 전기영동으로 HumTH01, HumTPOX, HumCSF1PO 및 Amelogenin을 동시에 분석할 수 있었다.

A simple and rapid procedure, called FoLT-PCR(Formamide Low Temperature-Polymerase Chain Reaction) was applied to amplifying DNA directly from various forensic biological evidences including human blood, saliva, hair root, or semen without any DNA preparative steps. We added washing step with non-ionic detergent, 1% Triton X-100, and used Taq DNA polymerase instead of Tth DNA polymerase to amplify 3 STR loci and gender allele simultaneouly. Optimal concentration of formamide and annealing temperature were determined empirically to 8%(v/v), and $48^{\circ}C$ respectively. We also compared this method with standard PCR.

키워드

참고문헌

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