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首页> 外文期刊>Nucleic acids research >PCR amplification of DNA microdissected from a single polytene chromosome band: a comparison with conventional microcloning
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PCR amplification of DNA microdissected from a single polytene chromosome band: a comparison with conventional microcloning

机译：从单个多染色体染色体条带上显微切割的DNA的PCR扩增：与常规微克隆的比较

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摘要

A novel alternative to microcloning for the production of region specific chromosomal DNA is described. In this method, ‘microamplification', single bands are dissected from polytene chromosomes and digested with Sau3A. Oligonucleotide adaptors are ligated to these fragments to provide convenient priming sites for polymerase chain reaction amplification. In this way, as much as 1μg of DNA can be amplified from a single band. Probes made from PCR amplified DNA from two such dissections have been used to probe cloned DNA form a 100kb chromosome walk. Whereas conventional microcloning has generated cloned EcoRI fragments corresponding to 3–4kb of the walk, the PCR probes cover greater than 90% of this chromosomal region. Thus microamplification is significantly more effective than microcloning in providing probes for establishing chromosomal walks.
机译：描述了用于产生区域特异性染色体DNA的微克隆的新型替代方法。在这种“微扩增”方法中，从多条染色体上切出单条带，并用Sau3A消化。将寡核苷酸衔接子连接至这些片段，以提供用于聚合酶链反应扩增的方便的引发位点。这样，从一个条带可以扩增多达1μg的DNA。由来自两个此类解剖结构的PCR扩增DNA制成的探针已用于探针从100kb染色体步移中克隆出的DNA。传统的微克隆产生了对应于步行3–4kb的克隆EcoRI片段，而PCR探针覆盖了该染色体区域的90％以上。因此，微扩增在提供用于建立染色体步移的探针方面比微克隆显着更有效。

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