ENHANCING YIELD OF LOW COPY NUMBER PLANT BASED VECTOR FROM E.COLI CELLS FOR GENE CLONING

Plasmid DNA is an important role play for gene cloning and gene expression analysis in E.coli cells. But some of the plasmids are very low copy number so its isolation is also difficult due to its low copy number. So in this study we used different types of media (broths) with more volume, area, specific growth temperature, time of incubation and quality of inoculum. We used five different types of broth for enhancing the yield of low copy number plasmids such as LB, LB24, SB, SOC and 2xYT media. In these five media the differences is low and high concentration of yeast extract and tryptone and in SOC media some different salts and glucose are additional. Comparison of these five media for enhancing the yield of low copy number plasmid DNA, SOC broth optical density is more comparative to other media. In SOC media plasmid culture OD and isolated plasmid DNA concentration both are directly proportional to each other. For enhancing the yield of low copy number plasmid DNA in E.coli cells these following factors are very important role play, quality of inoculum,size of inoculum specific incubation temperature, area of shaking vessel, incubation time, required concentration of antibiotics, and also concentration of tryptone, yeast extract and additional salts and glucose. One another important factor for enhancing the yield of low copy number plasmid used manual method (alkaline lysis method) for isolation of low copy number plasmid DNA and its purification also effects the yield of plasmid DNA.