LB

The medium used to grow E. Coli in is called LB, short for Lysogeny Broth. This contains nutrients that the bacteria likes, so that it grows well in it. A typical LB solution is made up of the following ingredients:

• tryptone
• yeast extract
• NaCl

An idea is to replace the above with the following ingredients, respectively:

• milk powder
• marmite
• table salt

(well, table salt is just the common name for NaCl anyway)

Arabinose

Arabinose is the promoter used in our DNA that makes the bacteria start expressing the fluorescent protein – in a way, switching on the fluorescent protein gene. According to [del Alamo], red wine contains arabinose in concentration levels that are sufficient for our needs. One needs to evaporate the alcohol from the wine first, as that would kill the bacteria.

Agar

Bacteria is grown on an agar gel for convenience, which is coincidentally used in East-Asian cuisine as well. So one just needs to get to a proper food store to get some.

Centrifuge

The process of creating competent cells involves centrifuging the sample, so as to collect all the cells together, and that one can get rid of the liquids around it. But bacteria cells can also be collected using filters aimed at creating sterile solutions, which in fact trap the bacteria. Filters of 20μm or 45μm are sufficient at trapping bacteria.

Need to do some further investigation though on where to get such filters, like in a pharmacy, for example.

Maarten de Smit just wrote me that there’s a way this can happen in a natural setting, by using water that is high on Calcium, according to [Baur]. Imagine – DNA transformation in a bottle of still mineral water

Or would you prefer sparkling?

Looking at the pGLO transformation manual (download the complete manual and the quick-guide from here, need to register first, which is free), we need to find kitchen-grade equivalents for the following tools, that are not included in the BIO-RAD kit:

• a way to create a precisely 42 °C water bath
• a way to create a 37 °C incubator (but this is actually not that important)
• micro-wave oven – well, this is easy

The following are included in the kit, but it would be nice if we could come up with a protocol that do not need these materials, but can be replaced with commercially available stuff:

• sterile pipets, with a resolution of 100μl and 250μl
• sterile innoculation loops
• sterile small test tubes
• sterile plates
• sterile water or sterile solution of CaCl₂

My gut feeling is that the above list is available in a regular pharmacy, one way or another. What seems to be really hard to get are:

• pGLO DNA plasmid
• competent E.Coli bacteria
• LB nutrient agar