WELCOME TO DIYBio.to!

We are a group of Bio-Enthusiasts and everything nice!

Ongoing Projects

Low Resource Protein Purification for Crowd-sourced Enzymes

The main project is the facile purification of DNA manipulating enzymes through fusing such enzymes to elastin-like polypeptides (ELPs). ELPs undergo a reversible phase transition in response to NaCl concentrations, in that raising the NaCl concentration of an ELP solution will cause the ELPs to aggregate. These aggregates can easily be separated from a complex mixture via centrifugation or microfiltration, and re-solubilized by lowering the NaCl concentration; a process termed ‘inverse transition cycling’ (ITC). Addition of a responsive self-cleaving peptide in between the protein of interest and the ELP, followed by another round of ITC, results in the purification of functional proteins.

We have two goals for this project:

1. To improve the ITC method by addition of a secretion tag to the ELP fusion proteins, precluding the need for cell lysis, as well as enabling the possibility of an efficient continuous production system.
2. Automation of the entire process through collaboration with computer scientists and roboticists which share our community maker-space.

The vision of our final product is a robot that produces functional workhorse enzymes on the bench top. People can create novel constructs and upload the sequence online, where others can download it and create in-house cell lines for the enzymes they need, which would all be purified by the robot. This would lower the burden of cost for lab work, lowering the bar of entry for entrepreneurs and innovators in the synthetic biology field.



Figure 1. The fusion protein. The protein-of-interest is green, the responsive self-cleaving peptide is yellow, and the ELP is red.


Figure 2. Protein-of-interest is green square (thioredoxin), a responsive self-cleaving peptide is the yellow oval, ELP is the yellow zig-zag line, other shapes are unwanted proteins in the mixture.

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Ge, X., Trabbic-Carlson, Chilkoti A., Filipe C.D.M. (2006). Purification of an elastin-like fusion protein bymicrofiltration. Biotechnology and Bioengineering, 95, 424-432.
Ge, X., Trabbic-Carlson, Chilkoti A., Filipe C.D.M. (2006). Simultaneous phase transition of ELP tagged molecules and free ELP: An efficient and reversible capture system. Biomacromolecules, 7, 2475-2478.

DIY Algae Photobioreactor

A small aquarium pump, some tubing, plastic bottles and hot glue are all that’s needed to make a photobioreactor. Our set-up is on display because it’s green, it bubbles and it looks cool as well as to show that scientific experiment is not an exotic affair, it’s within reach.


Schematic:

Experiment 1: Unprocessed Algae as fuel source
Step 1: Dehydrate Algae.
Step 2: Burn it

Related to Alage Biofuel Harvesting System

Future Projects

Meat Speciation

The current community project is species typing in commercial meat. DNA is extracted from the meat sample and species-specific primers are used to amplify DNA in the sample via the polymerase-chain-reaction (PCR). Analysis of the resulting DNA fragments shows which animal species were present in the meat.

Algae Biofuel Harvesting System

A major obstacle in the mass utilization of algae as sources of renewable fuel is the high cost of extracting the oil from inside the cells which results in destruction of the cell. To sidestep this obstacle, we would like to use Botryococcus braunii, an algae that lives in colonies and secretes oil into a communal pool which lies outside the cells. Our goal is to develop methods to extract the oil from the colonies with minimal damage to the cells, such that a continuous ‘milking’ process is achieved. The first step is acquiring the Botryococcus braunii and cultivating it continuously in the lab, and then comes the fun part, experimenting with oil extraction techniques. People are welcome to come up with their own ideas and implement them in the lab. Get your thinking caps on and we’ll see you in the lab! If you or someone you know can donate some Botryococcus braunii, please contact us! For now we are fundraising for $250 to purchase a strain.

Past Projects

Mini Maker Faire 2014- E. coli Art

At the mini maker last year we prepared some differently colored E. coli with Synbiota and their Rainbow kit and gave members of the public a chance to design their own art pieces with it!

Resouces

Downloads

A 3-D printable PCR Rack


Download:
1.5ml Centrifuge Tube Rack
0.2ml PCR Tube Rack