Difference between revisions of "Team:UCopenhagen/InterLab"

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Revision as of 13:57, 28 October 2017

I N T E R L A B

Introduction

We participated in InterLitab, as we want to be contribute to the scientific progress made through this globe spanning project. In InterLab, 6 test devices are inserted in E.coli D5 α, and the growth and fluorescence is measured.

We used the following plasmids provided by iGEM HQ to transform E.coli:

  • Positive control
  • Negative control
  • Test Device 1: J23101+I13504
  • Test Device 2: J23106+I13504
  • Test Device 3: J23117+I13504
  • Test Device 4: J23101.BCD2.E0040.B0015
  • Test Device 5: J23106.BCD2.E0040.B0015
  • Test Device 6: J23117.BCD2.E0040.B0015

Calibrations

Before our measurements began, we performed some calibrations: First an OD600 reference point for our plate reader, performed with LUDOX according to the protocol. Here we found a correction factor which can be used to calculate OD from measured absorbance. Our correction fator is 3.11.

Secondly we made a fluorescence standard curve with a serial dilution of fluorescein (figure 1). We used the lower 5 data points to calculate a mean µM fluorescein pr a.u. We chose to use the lower concentration range due to two factors: 1) Linearity is better for the lower fluorescein concentrations, and 2) our measured data has a maximum fluorescence of 500, which makes it more important to have a good fit in the lower range.



Figure 1. - Standard curve of fluorescein fluorescence. Fluorescence in arbitraty units (a.u.), fluorescein concentration in µM.

Cell measurements

Two colonies from each transformation were picked, and grown in foil-covered 50 ml falcon tubes over night (18 hours).

Preparation OD was measured, and a dilution was calculated to achieve an OD of 0.02. Dilution calculations can be found in the table next to this. Here we used the calculated correction factor from our initial abs/OD calibration. From the absorption measurements taken at 0 hours, we see indications of pipetting errors, as the OD600 (average) ranges from 0.015 to 0.6 (table 1).

Title
Abs600 OD600 before Volume (ml) added to 12 ml media OD600 after dilution (0 hours)
Sources: Wikipedia & Box Office Mojo. Data is current as of August 25, 2016.
Frozen 2013 Disney $1,287,000,000 $400,738,009 $875,742,326 $150,000,000
Minions 2015 Universal $1,159,398,397 $336,045,770 $823,352,627 $74,000,000
Toy Story 3 2010 Disney Pixar $1,066,969,703 $415,004,880 $651,964,823 $200,000,000
Zootopia 2016 Disney $1,023,227,498 $341,268,248 $681,959,250 $150,000,000
Despicable Me 2 2013 Universal $970,761,885 $368,061,265 $602,700,620 $76,000,000
The Lion King 1994 Disney $987,483,777 $422,783,777 $564,700,000 $45,000,000
Finding Nemo 2003 Pixar $936,743,261 $380,843,261 $555,900,000 $94,000,000
Shrek 2 2004 Dreamworks $919,838,758 $441,226,247 $478,612,511 $150,000,000
Finding Dory 2016 Disney Pixar $916,221,557 $478,714,390 $437,507,167 $250,000,000
Ice Age: Dawn of the Dinosaurs 2009 Fox $886,686,817 $196,573,705 $690,113,112 $90,000,000

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