What equipment do I need for a bacterial experiment?

Question

I am doing an experiment concerning evolution with bacteria and the antibiotic ampicillin. The idea is to see if, using small doses, the bacteria will develope a resistance to the antibiotic over time. What I need is a ligitimate list of materials and equipment that I will need to conduct this experiment. Also, if you think this will work or not, I could use some opinions. Thank you.

Answer 1

The list above is pretty good.

Your best chance of getting resistant colonies is to first generate bacteria which can survive a very small amout of amp, and then crank up the dosage.  A pre-made disk might not allow you to do this, so you may want to stick with solution, that you can add to the media in the desired concentration.

Also, you may want some kind of UV chamber, to mutate the bacteria faster.  Waiting for a random mutation might take longer than you have for your experiment.

Answer 2

Cultures of bacteria to be tested, gram positive and/or gram negative

Sterile culture media in petri dishes (Nutrient agar, tryptic soy agar, or Mueller-Hinton agar)

Sterile swabs

Incubator (or something to keep the temperature near body temperature)

Ampicillin solution or ampicillin disks

Forceps or Tweezers

Isopropyl alcohol (for flaming tweezers to sterilize the tweezers)

Plastic bucket and bleach to disinfect plates, swabs and other materials contaminated with the bacterial cultures.

Clorox Wipes or some other surface disinfectant to remove possible contaminant before and after performing the inoculations of bacteria onto the plates.

A good lab manual demonstrating sterile techniques and procedures

A microbiologist to supervise your experiment

You can find all these materials from Wards Scientific (www.wardsci.com)  or Carolina Biological Supply (www.carolina.com).  

You will need to work out a protocol for doing the work.  The use of the ampicillin impregnated disks will produce a clear zone where there is not visible bacterial growth.  If any of the cells are resistant, they will grow in the inhibition zone and form a colony.  You can sample that colony into a fresh dish and place a new ampicillin disk on the surface of the plate.  If the isolated culture is resistant, the zone of inhibition may be smaller or not present.

If you can't get the ampicilin you could do much the same exercise with sterile paper disks and antimicrobial soap.

Answer 3

A hazmat suit would be excellent!