"
Team:Evry/Notebook/w1
From 2013.igem.org
(Difference between revisions)
| Line 11: | Line 11: | ||
The chemically competent strains chosen are as follows:<br/> | The chemically competent strains chosen are as follows:<br/> | ||
<br/> | <br/> | ||
| - | |||
BL21<br/> | BL21<br/> | ||
DH5α<br/> | DH5α<br/> | ||
| - | TOP10<br/ | + | TOP10<br/> |
<br/> | <br/> | ||
| - | |||
<h2>Thursday, 20th June</h2> | <h2>Thursday, 20th June</h2> | ||
| + | <br/> | ||
| + | <b>Protocol for competent cells</b><br/> | ||
| + | <br/> | ||
| + | Inoculation of the bacteria:<br/> | ||
| + | For 200 ml LB medium, add 400 µL of strain sample (BL21 or DH5α).<br/> | ||
| + | However, for the TOP10 strain, we decided to make 400 mL LB medium with 800 µL of strain sample. In fact, TOP10 is a cloning strain and we will use it in our initial steps. | ||
| + | |||
Revision as of 14:54, 28 July 2013
Week 1: 17th June - 23rd June
Tuesday, 19th June
As a start for the labwork, we first decided to make competent cells. As a consequence, we grew from glycerol samples three E. coli strains in 2 mL of LB medium.The chemically competent strains chosen are as follows:
BL21
DH5α
TOP10
Thursday, 20th June
Protocol for competent cells
Inoculation of the bacteria:
For 200 ml LB medium, add 400 µL of strain sample (BL21 or DH5α).
However, for the TOP10 strain, we decided to make 400 mL LB medium with 800 µL of strain sample. In fact, TOP10 is a cloning strain and we will use it in our initial steps.
Friday, 21st June
The tree following E. coli strains have been prepared to be chemically competent: BL21, DH5α and TOP10.
To check the quality of our work as well as their competence, each strain has been plated on LB medium with ampicillin, kanamycin or chloramphenicol. We also transformed our bacteria with a random plasmid and plated them on LB medium with ampicillin only. We incubated our petri dishes for the week end at 30°c
