• 1 Shot glass
• Several Drops of Dish Soap – must contain sodium laurel sulfate
• 1 pinch of salt
• Contact lens solution OR meat tenderizer OR pineapple juice (optional)
• Ice-cold 120+ proof liquor (that’s 60% plus – Bacardi 151 should be sufficient)

-Rub the inside of your cheeks and rub your tongue around. Try and fill at least 1/4 of the glass with spit. This is harder than it sounds!  Think about bacon.

Spit contains cells from your mouth and cells contain DNA.   This is the same DNA that is in every cell in your body (except for those pesky microbes that are all over the place).

- Mix it around gently

Dish soap causes the cells in your spit to lyse (aka die, break open, explode).  Once they lyse, the DNA is released into the spit, just waiting for us to find it.

All three of those ingredients contain protease – enzymes which break down proteins.  Since we just lysed the cells, everything in them is floating around, including proteins.  To help extract the DNA we will break down some of these proteins using protease.

- Give it a gentle stir

DNA is a negatively charged molecule.  To help it precipitate we add salt (which forms Na+ and Cl- ions in water).  The Sodium (Na+ ions) will be attracted to the DNA.

This is important because we are trying to gather a large amount of DNA in one place.  If they are negatively charged they will repel each other.  If the negative charge is matched by a positive charge (the Sodium ions) it will make it much easier for the DNA to gather together.

-You must be VERY careful in this step to pour the alcohol on TOP of the spit and not let it mix.  You can do this by pouring it down the side of the glass very gently and slowly.  Note: you must have steady hands, so no drinking the alcohol beforehand.

DNA is very insoluble in alcohol, so the DNA at the surface of the spit precipitates out.  As this happens, DNA deeper in the spit solution gets drawn up and also precipitates.

- In the alcohol you should see a white cloudy mass, this is your DNA.  Apologies, you cannot actually see the helix.

You’re done!

Reference: http://www.instructables.com/id/5-minute-DNA-Extraction-in-a-Shot-Glass/

To achieve  our objective we have divided the project into 6 tracks that will be pursued in parallel:

1. Biofilm- observe Staphylococcus aureus biofilm growth under lab conditions so we can characterize effectiveness of our engineered phage

2. DspB- extract the Dispersin B gene from A. pleuropneumoniae  so we can integrate into the phage genome

3. Phage- develop standard protocol to manipulate the phage genome in a simple and well-defined manner

4. Quorum- characterize the existing P2 biobrick part

5. Modelling- model the interaction between s. aureus biofilm, phage, and relevant molecules in the system, to supplement our wet lab experiment

6. Human Practices- gather feedback from public opinion of synthetic biology

In the meantime, this Wednesday will be an Ideas Discussion meeting, so please come prepared to discuss the ideas put forth thus far. If possible, some faculty members will be present at the meeting as well to go through the ideas as well. Please bring your laptops if you can (so that you can access the wiki while at it).

Other than that, keep checking back for the latest research articles on Synthetic Biology!