IGEM:Cambridge/2008/Notebook/Turing Pattern Formation/2008/08/11: Difference between revisions

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|style="background-color: #EEE"|[[Image:IgemCam.jpg|200px]]<span style="font-size:22px;"> Turing Pattern Formation</span>
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==Erytromycin Experiment==
- New preparation of Ery : 0.05g into 10mL of ETHANOL
-We plated again our plates (same than 08/08/08)
==Amylase screening experiment==
- New method : add 2g of starch powder into 200mL of Agar, shake, pipette to plate (and avoid bubbles)
- Same plates than 08/08/08
==Test our stock of ECE171==
-Plasmid miniprep 9from the samples of 10mL and the one of 100mL)
-Nanodrop both DNA
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!  Vetor !! 260/280 !! μg/mL
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| ECE171 (10mL) || 1.73 || 136.3
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| ECE171(100mL) || 1.82 || 99.5
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- Double digest (EcoRI and PstI) with 10μL of DNA and 1h of incubation
- Samples have been frozen, they should be run onto a gel
==Control of Spc resistance of Bacillus==
- Spc50 plate with transformed  IA771 (ECE153, which is Spc resistant) and with IA771 9which should not be Spc resistant)
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__NOTOC__


==PCR Bacillus RBS==
==PCR Bacillus RBS==

Revision as of 08:23, 5 September 2008

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PCR Bacillus RBS

We created 2 different Bacillus RBS sites simply by primer annealing and extension. Each product includes an 8bp RBS plus the BioBrick prefix and suffix for a total of 54bp. The products were visualized on a 3.5% Agarose gel with bioline's hyperladder V.

RBSs is the consensus RBS sequence for Bacillus subtilis, and thus we believe it to be a very strong binding site - AAAGGAGG

RBSw has a 2bp modification from RBSs, which we believe will weaken it - AGAGGTGG


The amplification of RBSs yielded only one band on the gel. The product was purified using microclean and contained 14.8ng/uL after clean-up.

RBSw, however produced 2 bands on the gel. The correct size was gel-extracted and purified. After purification the yield was 12.3 ng/uL.

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