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Revision as of 09:06, 3 October 2018
Introduction
As Theodor Reik (Sigmund Freud disciple) said: “In science, like in all life fields, a slogan is imposed: security first”. Nowadays, safety is an essential requirement when creating new devices that humans will use, and Printeria is not different.
On the one hand biosafety describes the containment principles. technologies and practices that are implemented to prevent the unintentional exposure to pathogens and toxines, or their accidental release [1]. On the other hand biosecurity describes de protection, control and accountability for valuable biological materials within laboratories, inorder to prevent their unauthorized access, loss, theft, misuse, diversion or intentional release.
Next paragraphs describe how and why we implemented biosafety and biosecurity requirements in Printeria.
Lab safety
Safety design in Printeria
Lab Safety
This collaboration was born when Ismael (Imperial College iGEM team supervisor) came to the Univertitat Politécnica de Valencia to have a meeting with Diego Orzáez one of our instructors. Taking advantage of that situation we met with him and he explained us the Imperial College project called PixCell.
PixCell aims to get electronics and Synthetic Biology closer designing an innovative sistem where the culture is controlled by voltage.
Antes de poner en marcha el gran proyecto que es Printeria, tenemos que comprobar que las construcciones genéticas y los métodos de laboratorio que vamos a utilizar, funcionan en un entorno controlado y seguro como es el de un laboratorio. Printeria utiliza como chasis bacterias E.coli de riesgo biológico 1 provenientes de las cepas DB3.1, 10G y TOP10. Para las construcciones utilizamos biobricks y partes suministradas por IDT en Golden Gate. Este trabajo, es realizado solamente por personas con experiencia en el campo y con conocimiento sobre la seguridad del laboratorio, como es el caso de los Biotecnólogos y los Ingenieros Biomédicos. Protocolos seguridad de antemano escrita y guiada por los instructors Alejandro vignoni y Yadira Boada Acosta El laboratorio que utilizamos sigue la legislación vigente sobre el riesgo 1 (Ley 31/1995, RD 664/1997) y tiene para ello un autoclave () y una campana () con esterilización mediante UV. Además los resíduos generados por el laboratorio están minuciosamente controlados por el CGROM (centro generador de resíduos en la Oficina de Medioambiente) que facilitan los contenedores e intrucciones necesarias para la correcta clasificación de los resíduos.
Como medida de seguridad adicional, no se utiliza Bromuro de Etidio sino SYBR Safe, que tiene el mismo efecto sin efectos cancerígenos pero su uso es el mismo que con el BrEt en cuanto a protocolo.
Safety design in Printeria
Paris-Bettencourt team was right when they said : “Biosafety is an exciting design challenge, an essential enabling technology for synthetic biology, and a fundamental ethical obligation of all bioengineers”.
Safety Problem |
Solution |
|---|---|
Number of samples |
From 3 to 6 samples of reporter protein and 3 samples of blank |
Temperature |
37 ºC |
Shake |
Double Orbital. 01:00 (MM:SS) |
Absorbance. Optical Density (OD) |
Wavelenght at 600 nm emission |
Excitacion and emission scans |
The scans occur between two wavelength limit values. The established range will depend on the theoretical spectrum of the protein. |
Excitacion and emission wavelengths |
These values will depend on the range, and therefore on the spectrum. Values far from the theoretical peak lead to more attenuated fluorescence curves, and values very close to the peak can lead to overlap and error in reading the data. Therefore, a compromise must be reached between curve resolution and reading overlap. |
Gain (G) |
Normally the gain value G = 60, although for proteins with lower fluorescence, it is recommended that G takes higher values. |
Lista + Comité de ética. Riesgo y posibles soluciones.