TAILIEUCHUNG - Optimization of E. coli culture conditions for efficient DNA uptake by electroporation

Bacterial transformation is a significant breakthrough in the field of molecular biology for cloning purposes. A variety of chemical and physical methods are utilized for enhancing bacterial competence. The present study was designed, via the consolidation of chemical and physical methods, to improve competence of cells for high transformation efficiency up to 109 transformants/µg of DNA. | Turkish Journal of Biology Research Article Turk J Biol (2014) 38: 568-573 © TÜBİTAK doi: Optimization of E. coli culture conditions for efficient DNA uptake by electroporation Irfan AHMAD, Tehseen RUBBAB*, Farah DEEBA, Syed Muhammad Saqlan NAQVI Department of Biochemistry, Faculty of Biological Sciences, PMAS Arid Agriculture University, Rawalpindi, Pakistan Received: Accepted: Published Online: Printed: Abstract: Bacterial transformation is a significant breakthrough in the field of molecular biology for cloning purposes. A variety of chemical and physical methods are utilized for enhancing bacterial competence. The present study was designed, via the consolidation of chemical and physical methods, to improve competence of cells for high transformation efficiency up to 109 transformants/µg of DNA. LB and SOC media and their supplemented combinations with osmotic agents (sucrose and glycine) were used to prepare electrocompetent cells of E. coli DH5α at temperatures of 25 and 37 °C, which were then subjected to electroporation. The results showed that cells grown in SOC medium showed high growth rates and increased transformation efficiencies as compared to LB, while media supplemented with osmotic agents also showed higher transformation proficiency as compared to the controls. In all combinations tested, the highest transformation efficiencies were observed in the cells grown in SOC-SG at 25 °C, . × 109 cfu/µg of DNA. This study provides valuable knowledge for low-budget biotechnology labs to prepare competent cells with efficiency comparable to commercially available competent cells. Key words: Electroporation, optimization, culture conditions 1. Introduction In molecular biology, transformation is the uptake or introduction and expression of foreign DNA by living cells (Roychoudhry et al., 2009). Many bacteria have the ability to uptake .

• Sinh học
• Bacterial transformation
• Optimization of E
• coli
• Efficient DNA uptake
• Culture conditions
• Least significant difference
• Efficient protocol for Agrobacterium mediated transformation
• Gus gusplus gene into cassava plants
• Manihot esculenta Crantz
• Tumefaciens bacterial strains
• Three antibiotics kanamycin
• Environmental challenges
• Ligninolytic bacterial consortium
• The degradation of azo dye
• Electricity generation
• Dual chambered microbial fuel cell
• Bio transformation
• Antimicrobial resistance
• Minimum inhibitory concentration
• Plasmid transfermation
• Aquatic environment
• Spodoptera littoralis
• Biological activity
• Cotton leafworm
• Leaf dip method
• Salvia L
• species
• Ethnobotanical information
• Anti inflammatory activities
• In vivo experimental models
• Salt tolerance
• Saline water
• Oil crop
• Electrical conductivity
• Hybrid poplar
• Soil characteristics
• Mean annual height growth
• Food and agricultural organization
• Root to shoot ratio
• Soil salinity
• Chickpea Cicer arietinum L
• Ultimately higher yield