一种利用菌落直接PCR扩增DNA并用于测序的实验方法.通过对引物的设计和菌液浓度控制,使PCR反应后的内容物对测序干扰减到最小.与传统的测序过程比较,它省去了抽提模板DNA一步,节省了大量时间和实验成本.另外此方法可对BAC亚克隆库构建时由连接转化过程中导致的假阳性起筛选和鉴定作用.采用该法成功测定了籼稻(Oryza sativa indica)广陆矮4号的L3173号BAC DNA全长序列(约100 kb),GenBank登录号:AL512542.
A method of using colony PCR products to sequence directly by controlling primer design. Less time and cost than the normal way (without DNA extraction and sequencing prior purification). It can also detect the false insert fragments by electrophoreses of the PCR products. One BAC DNA of the Oryza sativa indica 4th chromosome has been sequenced by this method, the GenBank Accession number: AL512542.
唐晔盛,李英,朱静洁,胡欣,林志新,韩斌,洪国藩.菌落PCR在大规模基因组测序中的应用[J].生物化学与生物物理进展,2002,29(2):316-318
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