Team:UANL Mty-Mexico/Safety/id tags


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Identification (ID Tags)

Due to the recent increase in synthetic biology projects with great potential for direct application in the environment, industry, or any other activity that requires the release beyond the laboratory, it is necessary to take apart biosecurity measures to allow growth only in conditions specific and desired, we ensure the tracking of such agencies in the event that at some point these genetic technologies fail.

UANL team aware of this need, has designed a proposal for screening and identification of genetically modified organisms. This proposal is the design of small sequences flanking the prefix and suffix of the synthetic constructs, unique to each team participating in iGEM, and that can be detected by a single PCR.

Within the proposal also suggested that the primers required for the detection of genetically modified organisms of each institution participating in the iGEM be sent to the registry part so that they can be stored, and if necessary they can be easily distributed.

It is worth mentioning the importance and functions of these new "OMG ID Tag's" which are the:

  1. Comparison between Synthetic Organism and Wildlife Organism.
  2. Identification of synthetic organisms in nature are undesirable conditions which may affect the environment in which they are located.
  3. Possibility to contact the organization where that synthetic organism was created to gather information needed to expedite a contingency plan.

ID Tags - UANL

The design of our "ID Tag's" consist of a pair of sequences, one upstream of the prefix and suffix other downstream, using a set of nucleotides with a meaning for us.

Basically, the idea consists in creating the plasmid pSB1C3 personalized for each university that will be participating, they will have their ID tags such that with those designed primers just an specific site will be amplified, in that way each team will have two sequences that will recognize their constructions.

Figure 1. Prefix example of ID Tag. The codons in lowercase translate to form the spanish word "FLAMA" (Flame)
Figure 2. Suffix example of ID Tag. The codons in lowercase translate to form the spanish word "TIGRE" (Tiger)

PCR for detection

This option consists in a simple detection of primer’s design that will give us a specific size of band thanks to the arrangement of pieces. Although is the simplest way, it has issues being the main use of several primers for the detection of all the designed pieces. We are going to show the designed primers for this year pieces and information of interest like the melting temperature, the amplified piece and the size that will be created.

Figure 3. For the piece composed by pLac + 32°C RNA thermometer + GFP (LVA) [BBa_K1140002] we used the primers shown below

Figure 4. For the piece composed by pTet + 37°C RNA thermometer + RFP(LVA) [BBa_K1140002] we used the primers shown below
Figure 5. For the piece composed by pCons + 37°C RNA thermometer + LacI (LVA) [BBa_K1140002] we used the primers shown below

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