Team:NYU-AD/Interlab

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Interlab

Introduction

InterLab study is trying to resolve one of the greatest challenges in synthetic biology, which is that measurements of fluorescence usually can’t be compared because they are reported in different units or because different groups process data in different ways. Fluorescence is widely used as a proxy for promoter activity by expressing fluorescent proteins such as green fluorescent protein (GFP). While this is an indirect measurement, it provides a useful insight into expression levels and has the significant advantage that it can be monitored continuously without disrupting cells. This year iGEM has provided us with two protocols for measuring GFP fluorescence that will result in common, comparable units for teams to test out. The goal of the experiment is to determine how close can the numbers be when fluorescence is measured around the world.


InterLab Measurement Kit

This year teams were provided with the InterLab Measurement Kit which had to be stored at room temperature or 4C, It contained 7 tubes (5 with plasmid DNA and 3 with standard reagents):

  • Plasmid DNA (100 pg/uL in 10uL of Buffer EB)
    • Test Device 1: J23101.B0034.E0040.B0015 in pSB1C3
    • Test Device 2: J23106.B0034.E0040.B0015 in pSB1C3
    • Test Device 3: J23117.B0034.E0040.B0015 in pSB1C3
    • Positive Control Device: I20270 in pSB1C3. Also located in Kit Plate 3, well 8P
    • Negative Control Device: R0040 in pSB1C3. Also located in Kit Plate 2, well 6F
  • FITC Standard: one tube with dried down FITC for creating a FITC standard
  • LUDOX: one tube with 30% colloidal silica suspended in 1mL of water


Interlab Protocols

NYU-AD iGEM decided to follow the Plate Reader Protocol, which can be found here.

Our results can be found here.