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PREPARATION METHOD FOR IN-SITU HYBRIDIZATION PROBE

机译:原位杂交探针的制备方法

摘要

A preparation method for in-situ hybridization probes as follows: fragmenting objective DNAs, recovering 150-600 bp fragments, and after an enzyme modification, ligating, at intervals, the fragments with DNA adaptors containing restriction enzyme site sequences to large DNA loops and long chains; obtaining and labeling a large amount of DNAs in step A or B: A. isothermal amplifying, adding a single nucleotide substrate with a marker when amplifying, to obtain a DNA product with a marker; or B. isothermal amplifying, doping a single nucleotide substrate with a marker to the obtained product with a nick translation or random primer method, to obtain a DNA product with a marker; and digesting the DNA product with the marker by using corresponding restriction enzyme, to obtain in-situ hybridization probes with lengths of 150-600 bp. The method of the present invention accurately controls length range of the probes, reduces production cost, and improves product quality.
机译:原位杂交探针的制备方法如下:将目的DNA片段化,回收150-600 bp的片段,酶修饰后,将片段与含有限制性酶切位点序列的DNA衔接子间隔连接至大的DNA环和长链。链条在步骤A或步骤B中获得并标记大量DNA:A.等温扩增,扩增时在单核苷酸底物上添加标记,得到带有标记的DNA产物。或B.等温扩增,用切口平移法或随机引物法在单核苷酸底物上掺入标记物,得到DNA产物。用相应的限制性内切酶用标记物消化DNA产物,得到长度为150-600bp的原位杂交探针。本发明的方法精确地控制了探针的长度范围,降低了生产成本,并提高了产品质量。

著录项

  • 公开/公告号US2020248170A1

    专利类型

  • 公开/公告日2020-08-06

    原文格式PDF

  • 申请/专利权人 DIAGLOGIC BIOLABS (XIAMEN) CO. LTD.;

    申请/专利号US201816758283

  • 发明设计人 XIAOQUN JIANG;

    申请日2018-08-09

  • 分类号C12N15/10;C12Q1/6874;C12Q1/6827;

  • 国家 US

  • 入库时间 2022-08-21 11:20:37

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