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从黑腹果蝇多线染色体特定区域进行DNA的显微切割与克隆。

Microdissection and cloning of DNA from a specific region of Drosophila melanogaster polytene chromosomes.

作者信息

Scalenghe F, Turco E, Edström J E, Pirrotta V, Melli M

出版信息

Chromosoma. 1981;82(2):205-16. doi: 10.1007/BF00286105.

Abstract

Fragments from section 3 of the salivary gland X chromosome of D. melanogaster were dissected with a micromanipulator. The DNA was extracted, cut and ligated to a lambda vector in a volume of a few nanoliters in an oil chamber monitored through a microscope. From about 10 pg of DNA we obtained 80 recombinant clones, a sample of which were analysed and shown to contain Drosophila DNA which hybridises in situ to the region of section 3 of the X chromosome. With this technique we can isolate clones from any desired region as small as 200 kb from the euchromatic arms of polytene chromosomes.

摘要

用显微操纵器分离出黑腹果蝇唾液腺X染色体第3节段的片段。在通过显微镜监测的油室中,将DNA在几纳升的体积内进行提取、切割并连接到λ载体上。从大约10皮克的DNA中,我们获得了80个重组克隆,对其中一部分样本进行分析后发现,这些克隆含有能与X染色体第3节段区域进行原位杂交的果蝇DNA。利用这项技术,我们可以从多线染色体常染色质臂上分离出小至200 kb的任何所需区域的克隆。

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