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2005•Biotechnology(Faisalabad)Requires access

Comparative Study on Preparation and Storage of Competent Escherichia coli Cells

Deshun Shi

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Abstract

Objective:To establishing a high efficient protocol for preparation and storage of competent Escherichia coli cells.Methods:Three methods(SEM,TSS and CaCl_2 method) were compared for preparation of competent cells;Escherichia coli at different growth stages(optical density range from A_(600)=0.2 to 1.1) were choosed for preparation of competent cells bythe best method,meanwhile different conditions were used for storage of competent cells.Results:High transformation efficiency can be achieved by SEM at A_(600) of 0.36 or 0.58;cells stored with dimethylsulfoxide in -80℃ could keep consistently high transformation efficiency at least 40d.

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Objective:To establishing a high efficient protocol for preparation and storage of competent Escherichia coli cells.Methods:Three methods(SEM,TSS and CaCl_2 method) were compared for preparation of competent cells;Escherichia coli at different growth stages(optical density range from A_(600)=0.2 to 1.1) were choosed for preparation of competent cells bythe best method,meanwhile different conditions were used for storage of competent cells.Results:High transformation efficiency can be achieved by SEM at A_(600) of 0.36 or 0.58;cells stored with dimethylsulfoxide in -80℃ could keep consistently high transformation efficiency at least 40d.

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Available abstract

Objective:To establishing a high efficient protocol for preparation and storage of competent Escherichia coli cells.Methods:Three methods(SEM,TSS and CaCl_2 method) were compared for preparation of competent cells;Escherichia coli at different growth stages(optical density range from A_(600)=0.2 to 1.1) were choosed for preparation of competent cells bythe best method,meanwhile different conditions were used for storage of competent cells.Results:High transformation efficiency can be achieved by SEM at A_(600) of 0.36 or 0.58;cells stored with dimethylsulfoxide in -80℃ could keep consistently high transformation efficiency at least 40d.

Key concepts: Escherichia coli, Transformation (genetics), Chemistry, Food science, Biology, Biochemistry, Gene

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