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Team:TUDelft/14 July 2009
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='''14th July'''= | ='''14th July'''= | ||
| - | =='''Lab work'''== | + | =='''Lab work: Sriram & Orr'''== |
| - | Using | + | |
| - | + | Using 650 ul of competent cells and the transformation protocol, we inserted 13 biobricks for delay into those competent cells and grew them in a solid media to increase the amount of biobricks we will have. | |
| + | |||
| + | Diluted the 13 biobricks with 15 ul RNase free water and stored in -20°C freezer. | ||
| + | |||
| + | Standard transformation procedure: | ||
| + | |||
| + | Remove competent cells from -80, let thaw for 10 min on ice and aliquot in 50 ul amounts.<br> | ||
| + | add 2-5 ul of vector, usually in H2O, to 50 ul cells, no mixing by pipet due to shear induction.<br> | ||
| + | keep on ice for 20 minutes (vector spreading through volume).<br> | ||
| + | heat shock (42°C) for 45 seconds.<br> | ||
| + | keep on ice for 2 minutes.<br> | ||
| + | add 200 ul SOC, put on 37°C for 1 hour or longer with agitation (160 rpm).<br> | ||
| + | plate out 250 ul on appropriate antibiotics.<br> | ||
| + | |||
| + | Prepared 1 litre of LB agar medium and sent to Kluyver lab's kitchen. | ||
Revision as of 09:32, 15 July 2009
14th July
Lab work: Sriram & Orr
Using 650 ul of competent cells and the transformation protocol, we inserted 13 biobricks for delay into those competent cells and grew them in a solid media to increase the amount of biobricks we will have.
Diluted the 13 biobricks with 15 ul RNase free water and stored in -20°C freezer.
Standard transformation procedure:
Remove competent cells from -80, let thaw for 10 min on ice and aliquot in 50 ul amounts.
add 2-5 ul of vector, usually in H2O, to 50 ul cells, no mixing by pipet due to shear induction.
keep on ice for 20 minutes (vector spreading through volume).
heat shock (42°C) for 45 seconds.
keep on ice for 2 minutes.
add 200 ul SOC, put on 37°C for 1 hour or longer with agitation (160 rpm).
plate out 250 ul on appropriate antibiotics.
Prepared 1 litre of LB agar medium and sent to Kluyver lab's kitchen.
Weenink
Brought beaker of eppendorf tubes to autoclaving
