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Team:Cambridge/Lab book/Week 14
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Colonies grew. However colony PCR using the sequencing primers revealed them to be transformed with recircularised backbone. [[File:Unsuccessfulcolonypcr.jpg|300px|center|thumb| An amplicon of ~300 bp indicates no insert.]] | Colonies grew. However colony PCR using the sequencing primers revealed them to be transformed with recircularised backbone. [[File:Unsuccessfulcolonypcr.jpg|300px|center|thumb| An amplicon of ~300 bp indicates no insert.]] | ||
| - | The digestions and ligations were reattempted. | + | The digestions and ligations were reattempted, this time treating the backbone with alkaline phosphatase in order to reduce or eliminate self-ligation. |
'''Instrumentation''' | '''Instrumentation''' | ||
Revision as of 03:08, 27 September 2012
| Week: | 3 | 4 | 5 | 6 | 7 | 8 | 9 | 10 | 11 | 12 | 13 | 14 |
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Contents |
Monday (24/09/12)
Ratiometrica - Lux
Colonies grew. However colony PCR using the sequencing primers revealed them to be transformed with recircularised backbone.
The digestions and ligations were reattempted, this time treating the backbone with alkaline phosphatase in order to reduce or eliminate self-ligation.
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.
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.
Wednesday (26/09/12)
Thursday (27/09/12)
Friday (28/09/12)
