The present invention relates to an improved and efficient method for simultaneous monitoring of abundance of individual mutants of a microbe in mixed populations where
insertion of a known transposon in the
genome of a microbe such that each
mutant caries a single transposon
insertion, isolating the
genomic DNA of the mixed
population of mutants, fragmenting the same with a frequently
cutting restriction enzyme, ligating a double standard adapter to the
genomic DNA fragments, amplifying the
DNA fragments adjoining transposon insertions thereby generating a set of
DNA fragments corresponding only to mutated genes resolving the said amplified
DNA fragments followed by comparing the intensity of DNA fragments obtained from
population of mutants before selection with that obtained from the
population subjected to selection and finally sequencing the DNA fragments that change in abundance to identify the mutated genes.