Rapid construction of sequencing templates by random insertion of antibiotic resistance genes |
| |
Authors: | Nabirochkina Elena N Georgieva Sofia G Krasnov Aleksey N Soldatov Aleksey V |
| |
Affiliation: | Institute of Gene Biology, RAS Moscow, Russia. |
| |
Abstract: | We describe a novel and handy method for generating a population of templates for sequencing. The method is based on the random insertion of antibiotic resistance gene in plasmid DNA digested by DNase I. The advantages of this approach are the small quantity of DNA necessary for mutagenesis and the complete independence from the restriction map of the plasmid. DNase I digestion provides a random distribution of the insertions, while antibiotic selection provides low background. We also present a convenient PCR-based procedure for the analysis and ordering of obtained insertion mutants. |
| |
Keywords: | |
本文献已被 PubMed 等数据库收录! |