Team:Cambridge/Lab book/Week 14

Monday (24/09/12)

Ratiometrica - Lux

An amplicon of ~300 bp indicates no insert.

The digestions and ligations were reattempted.

Instrumentation

The sensitivity of the sensor concerning light intensities was tested using a dilution series of luciferase-producing bacteria. 20ml Cultures were grown overnight from single colonies. The cultures were induced with 40ul of 1.5M arabinose (for a final concentration of 3mM). Cultures were left for 2 1/2 hours for full induction. Subsequently, a culture was pelleted and resuspended in 4ml LB. Doubling dilutions, of volume 2ml, were made from this concentrate, down to 1/8th concentration. 1ml of each 2ml dilution was analysed in each cuvette, which was placed in the cuvette holder we made ourselves.

Raw sensor values(V) vs Concentration of bioluminescent E.coli- Concentrations on the x-axis are relative therefore an OD 600 value was also taken

Normalised sensor data using a dilution series of bioluminescent E.coli- Concentrations on the x-axis are relative therefore an OD 600 value was also taken

Normalised percentage of blue frequencies using the same dilution series of E.coli

Dilution series of E.coli from most concentrated to least

Tuesday (25/09/12)

Ratiometrica - Lux

a few colonies obtained on each plates. cultures made at the same time as preparing a colony PCR. Unfortunately, similar results were obtained to yesterday, showing no insert.

Instrumentation

The sensitivity of the sensor concerning different frequencies (colours) of light was tested as well. As can be seen below, measurements taken from orange and blue light yield values respectively above and below those from white light (our reference point). The data was taken using a constant intensity of light for each case (V.High and V.Low brightness). This was done with the aid of an Android phone and a specialised software application, called Color Flashlight, downloaded from the official Market.

Sensor data for different colours at different intensities

Wednesday (26/09/12)

Thursday (27/09/12)

Friday (28/09/12)