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    Method of inactivating ribonucleases at high temperature
    4.
    发明申请
    Method of inactivating ribonucleases at high temperature 审中-公开
    高温灭活核糖核酸酶的方法

    公开(公告)号：US20060211032A1

    公开(公告)日：2006-09-21

    申请号：US11406169

    申请日：2006-04-18

    申请人： Fen Huang ,  Christine Andrews ,  John Shultz

    发明人： Fen Huang ,  Christine Andrews ,  John Shultz

    IPC分类号： C12Q1/68 ,  C12P19/34 ,  C12N9/99

    CPC分类号： C12N15/1003 ,  C12N15/1096 ,  C12Q1/6806 ,  C12Q2527/127 ,  C12Q2527/101

    摘要： Methods for protecting RNA from RNase degradation and for inactivating RNases in solution are disclosed. The invention includes methods for protecting RNA during storage, for performing quantitative PCR reactions, or for preparing cDNA. The method includes using a combination of an RNase inhibitor in a solution containing or devoid of reducing agents and high heat to render RNases inactive.

    摘要翻译： 公开了用于保护RNA免受RNA酶降解和在溶液中灭活RNase的方法。 本发明包括保存RNA，进行定量PCR反应或制备cDNA的方法。 该方法包括在含有或不含还原剂和高热量的溶液中使用RNase抑制剂的组合以使RNase无活性。

    Methods for isolating nucleic acids using alkaline protease
    9.
    发明授权
    Methods for isolating nucleic acids using alkaline protease 失效
    使用碱性蛋白酶分离核酸的方法

    公开(公告)号：US5981235A

    公开(公告)日：1999-11-09

    申请号：US681922

    申请日：1996-07-29

    申请人： John Shultz ,  Craig E. Smith ,  Douglas R. Storts ,  Paula Brisco ,  Judy Frederiksen ,  Susanne Selman ,  Josephine Grosch

    发明人： John Shultz ,  Craig E. Smith ,  Douglas R. Storts ,  Paula Brisco ,  Judy Frederiksen ,  Susanne Selman ,  Josephine Grosch

    IPC分类号： C12P19/34 ,  C07H21/02 ,  C07H21/04 ,  C12N1/06 ,  C12N15/10 ,  C12N9/50 ,  C12N9/99

    CPC分类号： C12N15/1013 ,  C12N15/1003 ,  C12N15/1006

    摘要： Solutions containing nucleic acids are treated with an alkaline protease to digest proteins such as nucleases that degrade the nucleic acids. In the isolation of nucleic acids, a biological sample containing nucleic acids is suspended in a solution containing water, buffer and chelating agent, the pH of the solution is adjusted to at least about 10 by adding a solution of sodium hydroxide and anionic detergent, an alkaline protease is incubated in the solution until nucleases are degraded, the pH of the solution is lowered to reduce activity of the alkaline protease by adding a solution having a pH between 3.5 and 4.5 and the alkaline protease is heat inactivated. Lowering of the pH may produce a cloudy solution which is cleared by centrifuging. Nucleic acids are isolated from the cleared solution by alcohol precipitation, or by using paramagnetic particles or a resin matrix containing silica particles. A chaotropic salt can be used to reversibly bind DNA to the resin matrix.

    摘要翻译： 含有核酸的溶液用碱性蛋白酶处理以消化降解核酸的蛋白质如核酸酶。 在分离核酸时，将含有核酸的生物样品悬浮于含有水，缓冲剂和螯合剂的溶液中，通过加入氢氧化钠和阴离子洗涤剂溶液将溶液的pH调节至至少约10， 将碱性蛋白酶在溶液中孵育直到核酸酶降解，通过加入pH在3.5和4.5之间的碱性蛋白酶被热灭活，溶液的pH被降低以降低碱性蛋白酶的活性。 降低pH可能产生混浊溶液，通过离心清除。 通过醇沉淀或通过使用顺磁性颗粒或含有二氧化硅颗粒的树脂基质从澄清的溶液中分离出核酸。 离液盐可用于将DNA可逆地结合到树脂基质上。