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iGEM 2014 Measurement Interlab Study

Introduction

Please note: All Measurement Track teams are required to participate in the inter-lab study.

The goal of the inter-lab study is to obtain fluorescence data for three specific genetic devices expressing GFP from iGEM teams around the world. Can you measure fluorescence somewhere in your lab? Then this is the perfect study for you! Even if your lab or the organisms you work with mean that you can’t measure GFP from the specific devices, we want every team to be able to participate: email measurement@igem.org and we will work out an alternative.

All teams who participate in the inter-lab study will be acknowledged at the Giant Jamboree with a special award!

For any questions, contact measurement@igem.org.

Interlab Study Requirements

1. Each participating team must have an “Inter-lab Study” page on their wiki for easy reference for the Measurement Judges.
   1. All devices measured for this study should be listed on this page
   2. All protocols followed for this study should be linked/posted on this page
   3. Sequencing data for all measured devices should be posted on this page
   4. Note: A restriction map (a restriction digest run on a gel) confirming the correct size of the device will suffice if DNA sequencing is not possible

2. Each team must use the three BioBrick devices listed below in the “Three Required Devices” section.
   1. Note: Teams are encouraged to measure more devices (see “Extra Credit” section below), but the three required devices must be included for the Inter-Lab Study (unless an alternative set is required and negotiated via email to measurement@igem.org).

3. Each participating team must collect and submit fluorescence data for these three devices (the data will be submitted through the “Inter-Lab Study” form, see step 4).
   1. This data may be obtained by any means possible for the teams. Any instrument capable of measuring GFP is acceptable!
   2. Measurement data should be submitted in absolute units if possible. There are many ways this can be done, depending on your lab. If your lab cannot measure absolute units, relative units are acceptable.
      1. Hint: if you have access to a flow cytometer, absolute units can be measured by calibrating against standard fluorescent beads. A method for doing so is describe (and supported with online tools) at: https://synbiotools.bbn.com/
   3. Data must be measured in triplicate (i.e., three samples for each device).
      1. Note: The mean and standard deviation across the replicates must be included.
      2. All teams must fill in the “Inter-Lab Study” form where they will indicate the equipment used to measure the cells, describe how the data was collected, and list all controls used.
         

Required Devices

Important notes: The first device is already built and available in the distribution kit. The second and third devices must be built using the BioBricks standard protocol by the teams participating in this study.

1. Existing device: BBa_I20206 (J23101-B0032-E0040-B0015) in the pSB3K3 vector.
   1. Kit location
      1. Plate 4, Well 18A

2. New device to be built by the iGEM team: BBa_J23101 + BBa_E0240 (B0032-E0040-B0015), must be built in the pSB1C3 backbone
   1. Kit locations
      1. BBa_J23101 (called BBa_K823005 when in pSB1C3): Plate 1, Well 20K
      2. BBa_E0240 (in pSB1C3): Plate 2, Well 24B

3. New device to be built by the iGEM team: BBa_J23115 + BBa_E0240 (B0032-E0040-B0015), must be built in the pSB1C3 backbone
   1. Kit locations
      1. BBa_J23115 (called BBa_K823012 when in pSB1C3): Plate 1, Well 22I
      2. BBa_E0240 (in pSB1C3): Plate 2, Well 24B