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For the rapid sequencing of genomes the DNA is linearized automatically and at reduced costs with a mixture of proteases

机译:为了对基因组进行快速测序,可将DNA自动与蛋白酶混合物线性化并降低成本

摘要

Rapid sequencing of genomes by linearizing DNA, isolated genome DNA or un-isolated DNA sequence is placed on a longitudinal plate in a mixture of anaphase-, telophase-, G phase- or mitosis- or interphase chromosomes (10), broken down by proteases. It can also be laid on a rule or in a straight line along a rod or linearized. The DNA sequence is sequenced by laser scanning microscopy, weak ultra violet radiation absorbed by the DNA, screen tunnel microscopy, electron microscopy (EM), nuclear magnetic resonance (NMR) or other spectrographic analysis, circular dichroic (CD), and the like. The substrates can be moved on the long plate in opposite directions. Both ends of the genome sequence (7) are spliced with the free ends of the sequences immobilized in the recesses on the substrate, by the addition of DNA ligates and androstane triphosphate (ATP). As the movement of the substrate is in opposite directions, the sequences on the long plate are linearized on its surface or along the grooves. The sequences are immobilized on the substrate. The interphase are spliced by acting on the telome to release telome DNA sequences. The chromosome proteins are reduced or degraded in a mixture of proteases (11), without damage to the DNA. The surface of the long plate can be coated with a mixture of helicase such as 3'-5' E. Coli Rep helicase and ATP so that a double-strand DNA is divided into two single strands. The long plate can be elastic or rigid. During de condensation and linearizing the chromosome DNA is irradiated with bio photons, the photons are then detected by photo-sensors such as photo-multipliers. The detection signals are stored and/or processed. The long plate has the groove and/or recess or reagent glass. The long plate or only the recess can be heated to release hydrogen bridges or DNA-DNA interactions. They can also be released by the addition of a denaturing material. A number of fragments of genome or chromosome DNA sequences (7) can be sequenced on the long plate simultaneously, in a multi-parallel sequencing action, in an automatic process. The movement of the substrate along the long plate is controlled automatically.
机译:通过线性化DNA,分离的基因组DNA或未分离的DNA序列对基因组进行快速测序,将其放置在纵板中的后期相,端期相,G相或有丝分裂或中间相染色体的混合物中(10),被蛋白酶分解。也可以将其放置在尺子上或沿着杆沿直线放置或线性化。通过激光扫描显微镜,DNA吸收的弱紫外辐射,屏幕隧道显微镜,电子显微镜(EM),核磁共振(NMR)或其他光谱分析,圆二色性(CD)等对DNA序列进行测序。基板可以在长板上沿相反方向移动。通过添加DNA连接物和三磷酸雄甾烷(ATP),将基因组序列(7)的两端与固定在基板凹口中的序列的自由端剪接。由于基板的移动方向相反,因此长板上的序列在其表面或沿凹槽线性化。序列固定在底物上。通过作用于端粒以释放端粒DNA序列来剪接中间相。染色体蛋白在蛋白酶混合物中被还原或降解(11),而不会损坏DNA。长板的表面可以涂有解旋酶(例如3'-5'E. Coli Rep解旋酶和ATP)的混合物,以便将双链DNA分为两条单链。长板可以是弹性的或刚性的。在脱凝和线性化过程中,用生物光子辐照染色体DNA,然后由光传感器(例如光电倍增管)检测光子。检测信号被存储和/或处理。长板具有凹槽和/或凹槽或试剂玻璃。可以加热长板或仅凹槽,以释放氢桥或DNA-DNA相互作用。它们也可以通过添加变性材料来释放。许多基因组或染色体DNA序列片段(7)可以在长板上以自动测序的方式同时进行多平行测序。基板沿着长板的移动是自动控制的。

著录项

  • 公开/公告号DE19937512A1

    专利类型

  • 公开/公告日2001-02-15

    原文格式PDF

  • 申请/专利权人 CHERKASKY ALEXANDER;

    申请/专利号DE1999137512

  • 发明设计人 CHERKASKY ALEXANDER;

    申请日1999-08-09

  • 分类号C12Q1/68;C12M1/00;

  • 国家 DE

  • 入库时间 2022-08-22 01:10:22

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