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Team:UNIPV-Pavia/Notebook/Week1Jul
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| + | *Miniprep for the overnight cultures. | ||
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| + | *Digestion E-P for the purified plasmids (under the same conditions as the experiment of June 30th). | ||
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| + | *Gel results: | ||
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Revision as of 22:36, 2 July 2009
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Week from July 1st, to July 5th, 2009
Previous Week
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Next Week
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July, 1st
- Sequencing results for BOL1 BioBrick: sequence was correct!
- Miniprep for:
- A8 (for dilution/transformation)
- A9 (for dilution/transformation)
- We transformed 10 pg of A8 and A9 miniprepped DNA in TOP10, after a proper quantification with Nanodrop and dilution. We plated transformed bacteria and incubated them overnight at 37°C.
- We prepared 0.5 l of LB + Amp.
- We infected 5 ml of LB + Amp with 10 ul of T9002 glycerol stock. We incubated the culture overnight at 37°C, 220 rpm.
- We also picked a random colony from A10 plate and infected 1 ml of LB + Amp. We incubated this inoculum at 37°C, 220 rpm for 5 and 1/2 hours. Then we prepared a glycerol stock and re-filled the remaining 250 ul of culture with 5 ml of LB + Amp to grow an overnight culture (for sequencing).
Preparation of experiment with Tecan F200
- We diluted 1:1000 the cultures of:
- J23100
- J23101
- J23118
- A1
- A2
- A7
- E0240
- We incubated the diluted cultures at 37°C, 220 rpm for 3 hours.
Experiment with Tecan F200
- Description
- Purpose:
- Materials & Methods
- Protocol
- Results
July, 2nd
- Miniprep for A10 and T9002. We sent purified DNA to BMR Genomics for sequencing.
- A8(10pg) and A9(10pg) plates showed colonies!(a few, as expected) We picked 3 colonies for A8 plate and 3 colonies for A9 plate to infect 5 ml of LB + Amp. We incubated the six inocula at 37°C, 220 rpm overnight. Tomorrow they will be miniprepped and screened with digestion to check if the extra band had been lost!
Preparation of experiment with Tecan F200
- aTc stock preparation.
- We diluted 1:1000 the cultures of:
- A8
- A9 (X2)
- E0240
- We incubated the diluted cultures at 37°C, 220 rpm for 3 hours. After 2 hours, one of the two A9 culture was induced with 75 ng/ml aTc and incubated again under the same conditions as before.
Experiment with Tecan F200
- Description
- Purpose:
- Materials & Methods
- Protocol
- Results
July, 3rd
- Miniprep for the overnight cultures.
- Digestion E-P for the purified plasmids (under the same conditions as the experiment of June 30th).
- Gel results:
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Previous Week
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Next Week
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