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KR960701081A - 산프로테아제를 사용한 핵산의 제조(Preparation of Nucleic Acids Using Acid Protease) - Google Patents

산프로테아제를 사용한 핵산의 제조(Preparation of Nucleic Acids Using Acid Protease) Download PDF

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KR960701081A
KR960701081A KR1019950703298A KR19950703298A KR960701081A KR 960701081 A KR960701081 A KR 960701081A KR 1019950703298 A KR1019950703298 A KR 1019950703298A KR 19950703298 A KR19950703298 A KR 19950703298A KR 960701081 A KR960701081 A KR 960701081A
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biological sample
protease
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nucleic acid
exogenous
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다니엘 엘. 캐시앙
키요타다 누노무라
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다니엘 엘. 캐시앙
젠-프로브 인코포레이티드
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Publication of KR960701081A publication Critical patent/KR960701081A/ko

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Abstract

목적 핵산(들)을 분해시킬 수 있는 내인성 뉴클레아제가 불활성으로 되는 pH로 생물학적 시료를 산성화시키고, 상기 생물학적 시료를 상기 pH에서 활성인 외인성 산프로테아제와 접촉시키고, 생물학적 시료를 내인성 뉴클레아제 활성이 유의적이지 않은 수준으로 감소될 때까지 배양하고, 상기 생물학적 시료의 pH를 상기 외인성 프로테아제의 활성을 저하시키기에 충분한 pH로 상승시키는 단계를 포함하여 생물학적 시료중에 포함되어 있는 유용한 목적 핵산을 제조하는 방법이 기술되어 있다.

Description

산프로테아제를 사용한 핵산의 제조(Preparation of Nucleic Acids Using Acid Protease)
본 내용은 요부공개 건이므로 전문내용을 수록하지 않았음

Claims (18)

  1. 생물학전 시료를 목적 핵산(들)을 분해시킬 수 있는 내인성 뉴클레아제가 부활성화 되는 pH로 산성화시키고, 상기 생물학적 시료를 상기 pH에서 활성인 외인성 산프로테아제와 접촉시키고, 내인선 뉴클레아제 활성이 상당한 수준으로 감소될 때까지 상기 생물학적 시료를 배양하고, 상기 생물학적 시료의 pH를 상기 외인성 프로테아제의 활성을 저하시키기에 충분한 pH로 상승시키는 단계를 포함하는, 생물학적 시료중에 포함되어 유용한 목적 핵산의 제조 방법
  2. 제1항에 있어서, 상기 생물학적 시료가 감염성 질환 작(作因)을 함유할 수 있는 인간 생물학적 물질인 것인 방법
  3. 제1항에 있어서, 상기 생물학적 시료가 인간 세포질 핵산을 함유하는 세포인 것인 방법
  4. 제1항에 있어서, 상기 산프로테아제가 펩신인 것인 방법
  5. 제1항에 있어서, 상기 배양 단계 이후의 시료의 pH를 후소 사용에 적합한 수준 까지만 상승시키고, 외인성 프로테아제가 완전히 억제되거나 또는 불활성화되는 수준까지는 상승시키지 않는 것인 방법
  6. 제1항에 있어서, 상기 산성화 단계에서 pH를 1.0 내지 4.0으로 조정하는 것인 방법
  7. 제1항에 있어서, 상기 pH상승 단계에서 pH를 6.0 보다 높게 조졸하는 것인 방법
  8. 제1항에 있어서, 상기 pH상승 단계이후 상기 시료를 가열함으로써 산프로테아제의 불활성화를 촉진시키는 것인 방법
  9. 제1항에 있어서, 세정제를 시료에 가하여 다른 시료 성분들로 부터 목적 핵산의 방출을 촉진하는 방법
  10. 제1항에 있어서, 상기 배양 시간을 내인선 뉴클레아제 농도를 유의적이지 못한 수준으로 감속시키기에 필요한 시간보다 길게 하여 상기 핵산을 시료 성분들로 부터 단리시키는 것을 촉진시키는 것인 방법
  11. 별도의 격실내에 생물학적 시료의 pH를 pH 4.0 또는 그 미만으로 감소시키기에 적합한 산, 및 생물학적 시료 중의 세포질 물질을 분해하여 그 시료 중의 핵산을 단리시키고 존재할 수 있는 내인성 뉴클레아제를 불활성화시키기에 적합한 산프로테아제를 포함하는 키트
  12. 제11항에 있어서, 단백질 분해 단계 종결 이후 시료의 pH를 상승시키기에 적합한 염기를 별도의 결실내에 추가로 포함하는 키트
  13. 프로테아제 용액을 RNAㅇ제 흡착제가 상기 프로테아제 용액 중의 RNA제즌 흡수하고, 프로테아제는 흡수하지 않는 조건하에 RNA제 흡착제로 처리하고, 상기 흡착제를 상기 프로테아제로부터 단리비시튼 단계를 포함하는, 잔류 RNA제를 제거하기 위한 프로테아제의 정제 방법
  14. 제13항에 있어서, 상기 프로테아제가 펩신인 것인 방법
  15. 제2항에 있어서, 상기 인간 생물학적 물질이 조직 세포 또는 혈액 성분인 것인 방법
  16. 제3항에 있어서, 상기 세포가 인간 백혈구 또는 중앙 세포인 것인 방법
  17. 제1항에 있어서, 상기 생물학적 시료가 인간 혈청인 것인 방법
  18. 생물학적 시료를 목적 핵산(들)을 분해시킬 수 있는 내인성 뉴클레아제의 활성이 유의적이지 않은 수준으로 감소되는 pH로 산성화되고, 상기 생물학적 시료를 상기 pH에서 활성인 외인선 산프로테아제와 접촉시키고, 상기 생물학적 시료를 내인성 뉴클레아제의 활성이 유의적이지 않은 수준으로 비가역적으로 감소될 때까지 배양하고, 상기 생물학적 시료의 pH를 상기 외인성 프로테아제의 활성을 저하시키기에 충분한 pH로 상승시키는 단계를 포함하는, 생물학적 시료 중에 포함된 유용한 목적 핵산(들)의 제조 방법
    ※ 참고사항 : 최초출원 내용에 의하여 공개하는 것임
KR1019950703298A 1993-02-10 1994-02-07 산프로테아제를 사용한 핵산의 제조(Preparation of Nucleic Acids Using Acid Protease) KR960701081A (ko)

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US08/015,729 US5386024A (en) 1993-02-10 1993-02-10 Method to prepare nucleic acids from a biological sample using low pH and acid protease
US08/015,729 1993-02-10
PCT/US1994/001421 WO1994018238A1 (en) 1993-02-10 1994-02-07 Preparation of nucleic acids using acid protease

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KR960701081A true KR960701081A (ko) 1996-02-24

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US (2) US5386024A (ko)
EP (1) EP0611157B1 (ko)
JP (1) JP3477205B2 (ko)
KR (1) KR960701081A (ko)
AT (1) ATE238430T1 (ko)
AU (2) AU690895B2 (ko)
CA (1) CA2155744C (ko)
DE (1) DE69432540T2 (ko)
DK (1) DK0611157T3 (ko)
ES (1) ES2198411T3 (ko)
WO (1) WO1994018238A1 (ko)

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EP0611157A3 (en) 1994-12-28
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DE69432540D1 (de) 2003-05-28
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US5386024A (en) 1995-01-31
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