PLASMID SCREENING AT HIGH COLONY DENSITY
PLASMID SCREENING AT HIGH COLONY DENSITY
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DOI：
10.1016/0378-1119(80)90144-4
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发表时间：
1980-01-01
期刊：
影响因子：
3.5
通讯作者：
MESELSON, M
中科院分区：
文献类型：
--
作者：
HANAHAN, D;MESELSON, M
描述了一种通过高密度核酸杂交筛选含有重组质粒的细菌（包括大肠杆菌）菌落的方法，即，每150 mm直径平板100，000个菌落。小菌落在硝酸纤维素滤膜上建立，它们可以忠实地复制到其他滤膜上。氯霉素扩增可在筛选前原位进行。过滤器可以冷冻保存，以长期储存解冻后可以进一步复制的菌落。[This研究可用于重组DNA研究和基因工程]。
A procedure is described for screening bacterial (including Escherichia coli) colonies containing recombinant plasmids by nucleic acid hybridization at high density, i.e., at 100,000 colonies per 150 mm diameter plate. Small colonies are established on nitrocellulose filters from which they can be faithfully replicated to additional filters. Chloramphenicol amplification may be carried out in situ before screening. The filters may be kept frozen for long-term storage of colonies which may be further replicated after thawing. [This study had applicability to recombinant DNA research and genetic engineering].