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    • 6. 发明授权
    • Optical selection and collection of DNA fragments
    • DNA片段的光学选择和收集
    • US5879625A
    • 1999-03-09
    • US951955
    • 1997-10-17
    • Mary C. RoslaniecJohn C. MartinJames H. JettL. Scott Cram
    • Mary C. RoslaniecJohn C. MartinJames H. JettL. Scott Cram
    • C12N15/10C12N15/12C12Q1/68G01N15/14G01N1/14
    • G01N15/1459C12N15/1006C12Q1/68G01N2015/0038G01N2015/149Y10T436/143333
    • Optical selection and collection of DNA fragments. The present invention includes the optical selection and collection of large (>.mu.g) quantities of clonable, chromosome-specific DNA from a sample of chromosomes. Chromosome selection is based on selective, irreversible photoinactivation of unwanted chromosomal DNA. Although more general procedures may be envisioned, the invention is demonstrated by processing chromosomes in a conventional flow cytometry apparatus, but where no droplets are generated. All chromosomes in the sample are first stained with at least one fluorescent analytic dye and bonded to a photochemically active species which can render chromosomal DNA unclonable if activated. After passing through analyzing light beam(s), unwanted chromosomes are irradiated using light which is absorbed by the photochemically active species, thereby causing photoinactivation. As desired chromosomes pass this photoinactivation point, the inactivating light source is deflected by an optical modulator; hence, desired chromosomes are not photoinactivated and remain clonable. The selection and photoinactivation processes take place on a microsecond timescale. By eliminating droplet formation, chromosome selection rates 50 times greater than those possible with conventional chromosome sorters may be obtained. Thus, usable quantities of clonable DNA from any source thereof may be collected.
    • DNA片段的光学选择和收集。 本发明包括从染色体样品中大量(>μg)量的可克隆,染色体特异性DNA的光学选择和收集。 染色体选择基于不需要的染色体DNA的选择性不可逆光激活。 尽管可以设想更一般的程序,但是通过在常规流式细胞术装置中处理染色体但是不产生液滴来证明本发明。 样品中的所有染色体首先用至少一种荧光分析染料染色,并与光化学活性物质结合,如果激活可使染色体DNA不可克隆。 在通过分析光束之后,使用被光化学活性物质吸收的光照射不想要的染色体,从而引起光激活。 根据需要,染色体通过该光激活点,灭活光源被光学调制器偏转; 因此,期望的染色体不是光激活的并且保持克隆。 选择和光激活过程在微秒的时间尺度上进行。 通过消除液滴形成,可以获得比常规染色体分选机可能的染色体选择率高50倍的染色体选择率。 因此,可以收集来自其任何来源的可克隆DNA的可用量。