Academic journal article Iranian Journal of Public Health

Using 2x Concentration (200Rg/ml) of Ampicilin Is the Most Optimum Concentration for Growing Bacteria Regard to Plasmid Extraction

Academic journal article Iranian Journal of Public Health

Using 2x Concentration (200Rg/ml) of Ampicilin Is the Most Optimum Concentration for Growing Bacteria Regard to Plasmid Extraction

Article excerpt

Background: The aim of this study is to determine the best concentration of medium antibiotics for the best yield of plasmid extraction.

Methods: Escherichia coli (E.coli) strain Top 10 F' was used as host for Green Fluorescent Protein (GFP) expression vector encoding ampicillin resistance gene. E.coli Top 10 was grown in Luria-Bertani (LB) broth medium containing 50 µg/ml tetracycline under aerobic condition and then has subjected to transformation with the vector of interest. The trans- formed E.coli was then cultured in LB broth containing 100 µg/ml Ampicillin for 4 hours. Then 150 µl of this medium were added to 4.5 ml of LB broth medium containing differ- ent concentrations (0,100, 200 and 300 µg/ml) of ampicillin and cultured at 37°C with shaking at 250 rpm for 16 hours. The medium containing bacteria were washed with sterile PBS and analyzed with flowcytometry.

Results: The flow cytometric results showed that the popu- lation of GFP positive bacteria was grown in 200 µg/ml of ampicillin was higher than other bacteria grown in 0, 100 or 300 µg/ml of ampicillin. …

Search by... Author
Show... All Results Primary Sources Peer-reviewed

Oops!

An unknown error has occurred. Please click the button below to reload the page. If the problem persists, please try again in a little while.