Team:TUDelft/14 July 2009

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='''14th July'''=
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=='''Lab work'''==
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=='''Lab work: Sriram & Orr'''==
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Using some competent cells and the transformation protocol, we inserted a couple of biobricks into those competent cells and grew them in a solid media to increase the amount of biobricks we will have.
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Protocol to be inserted...  
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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.
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Diluted the 13 biobricks with 15 ul RNase free water and stored in -20°C freezer.
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Standard transformation procedure:
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Remove competent cells from -80, let thaw for 10 min on ice and aliquot in 50 ul amounts.<br>
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add 2-5 ul of vector, usually in H2O, to 50 ul cells, no mixing by pipet due to shear induction.<br>
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keep on ice for 20 minutes (vector spreading through volume).<br>
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heat shock (42°C) for 45 seconds.<br>
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keep on ice for 2 minutes.<br>
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add 200 ul SOC, put on 37°C for 1 hour or longer with agitation (160 rpm).<br>
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plate out 250 ul on appropriate antibiotics.<br>
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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