Difference between revisions of "Team:Aix-Marseille/Manipulation"
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E. coli being aerobic, the solution is stored in LB in a Erlenmeyer flask of 5 × volume, so 500 mL and incubate for 1 h. | E. coli being aerobic, the solution is stored in LB in a Erlenmeyer flask of 5 × volume, so 500 mL and incubate for 1 h. | ||
| − | + | Glycerolization of strains | |
The fact of putting the strains in glycerol will protect them from the cold, necessary when one will freeze at -80 ° C so that the cells do not die. | The fact of putting the strains in glycerol will protect them from the cold, necessary when one will freeze at -80 ° C so that the cells do not die. | ||
40% Glycerol in 1.8mL cryotubes → 0.9 of glycerol and 0.9 of strains. | 40% Glycerol in 1.8mL cryotubes → 0.9 of glycerol and 0.9 of strains. | ||
We will freeze the cells in exponential phases in order to make our cells competent, ie able to incorporate DNA | We will freeze the cells in exponential phases in order to make our cells competent, ie able to incorporate DNA | ||
Revision as of 13:09, 14 June 2017
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Day 1 : 06/06/2017
Cleaning the laboratory, then recovery and installation of the equipment. Starting culture of strains TG1 and DH5α. We launched precultures from cryotubes ( sampling with 1 rod and then deposition in an erlenmeyer containing LB media ). Incubation at 37 ° C.
Day 2 07/06/2017
Strains cultivation: Recovery of strains cultured the day before: DH5α and TG1 To check the concentrations, the OD is measured. For this purpose, the culture must be diluted because the optimum measurement of the apparatus is between 0.1 and 0.8. The cultures in the tank are diluted to 1/10 with distilled water. Therefore, TG1: 5.16 / DH5α: 5.21 To calculate the volume to be taken for our 100 mL we do: DO WantedDO Obtained before dilutionVolume → 0.15.16 100 2 mL E. coli being aerobic, the solution is stored in LB in a Erlenmeyer flask of 5 × volume, so 500 mL and incubate for 1 h.
Glycerolization of strains The fact of putting the strains in glycerol will protect them from the cold, necessary when one will freeze at -80 ° C so that the cells do not die. 40% Glycerol in 1.8mL cryotubes → 0.9 of glycerol and 0.9 of strains. We will freeze the cells in exponential phases in order to make our cells competent, ie able to incorporate DNA
