IGEM:Cambridge/2008/Notebook/Turing Pattern Formation/2008/08/11

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Revision as of 06:37, 5 September 2008



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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

Vetor 260/280 μg/mL
ECE171 (10mL) 1.73 136.3
ECE171(100mL) 1.82 99.5

- 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)



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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