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Electrocompetent cells: Difference between revisions
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*Resuspend in GYT: Resuspend in 1 ml ice cold GYT. This is best done by gently swirling rather pipetting or vortexing. | *Resuspend in GYT: Resuspend in 1 ml ice cold GYT. This is best done by gently swirling rather pipetting or vortexing. | ||
*Measure OD: Measure the OD-600 of a 1:100 dilution of the cell suspension. (In the cuvette, mix 0.99 mL water and 0.01 mL cell | *Measure OD: Measure the OD-600 of a 1:100 dilution of the cell suspension. (In the cuvette, mix 0.99 mL water and 0.01 mL cell suspension). | ||
''Note: The desired concentration is <math>2.5 \times 10^{11}</math> cells per mL, giving <math>1\times 10^{10}</math> cells per 40 <math>\mu</math>L. This corresponds to an OD-600 (after 100x dilution) of roughly 3.75. It is difficult to reach this value, but it is still important to know the concentration of cells to calculate efficiencies. | ''Note: The desired concentration is <math>2.5 \times 10^{11}</math> cells per mL, giving <math>1\times 10^{10}</math> cells per 40 <math>\mu</math>L. This corresponds to an OD-600 (after 100x dilution) of roughly 3.75. It is difficult to reach this value, but it is still important to know the concentration of cells to calculate efficiencies. | ||
