The lab so far.
Mar. 7th, 2004 07:29 pm![[personal profile]](https://www.dreamwidth.org/img/silk/identity/user.png){kind=small}
gemfyreI was actually in the mood to do work so I wrote out what I know of my experiment so far. Any suggestions, ideas, help would be great!! I seemed to come up with more questions than answers.
The story so far...
Ingredients
20 petri dishes
400mL nutrient agar
1 McCartney bottle
Streaking loop
Bunsen burner (always gotta have them)
Laminar flow hood (ooer)
Method
Pour agar into 20 petri dishes.
1st plating - 2 dishes
2nd plating - 8 dishes (2 for each colony)
3rd plating - 8 dishes (2 for each colony)
Collect sample from soil at contaminated site in Bellevue (find info about Bellevue fire).
Plate onto two dishes.
Incubate 1 week.
Take sample from four different colonies, plate on 2 dishes each. Incubate (how long??)
Repeat next week.
Now we (hopefully) have nice pure cultures.
Gram stain cultures. Find the gram negatives.
Test for growth on heavy metals. Maybe choose metal most likely to have contaminated original site (mercury?) What concentration??
Nutrient agar with metal included. Not exactly sure how to do this. Diluted dots on the plate? No idea what the original concentration of cells will be so no idea how much to dilute.
For the stuff that does grow. Grow a nice sample in nutrient broth. (perhaps keep metal concentration in the broth the same, yeah, I think that's a good idea.)
Conjugate with non-resistant bacteria (which one? E. coli?)
Test if conjugated bacteria grows (how will I know which cells are the originals and which are the receivers??)
THEN, get the original culture. Start growing it in plain broth. Take samples at time intervals (how long interval? 10 minutes? 20 minutes? What's the replication time of my original sample? I have no way of knowing.) Grow on plates with metal.
See what grows, how long does it take to lose resistance? Does it lose resistance at all?
After all that crap
Write the smegging lab report.
The story so far...
Ingredients
20 petri dishes
400mL nutrient agar
1 McCartney bottle
Streaking loop
Bunsen burner (always gotta have them)
Laminar flow hood (ooer)
Method
Pour agar into 20 petri dishes.
1st plating - 2 dishes
2nd plating - 8 dishes (2 for each colony)
3rd plating - 8 dishes (2 for each colony)
Collect sample from soil at contaminated site in Bellevue (find info about Bellevue fire).
Plate onto two dishes.
Incubate 1 week.
Take sample from four different colonies, plate on 2 dishes each. Incubate (how long??)
Repeat next week.
Now we (hopefully) have nice pure cultures.
Gram stain cultures. Find the gram negatives.
Test for growth on heavy metals. Maybe choose metal most likely to have contaminated original site (mercury?) What concentration??
Nutrient agar with metal included. Not exactly sure how to do this. Diluted dots on the plate? No idea what the original concentration of cells will be so no idea how much to dilute.
For the stuff that does grow. Grow a nice sample in nutrient broth. (perhaps keep metal concentration in the broth the same, yeah, I think that's a good idea.)
Conjugate with non-resistant bacteria (which one? E. coli?)
Test if conjugated bacteria grows (how will I know which cells are the originals and which are the receivers??)
THEN, get the original culture. Start growing it in plain broth. Take samples at time intervals (how long interval? 10 minutes? 20 minutes? What's the replication time of my original sample? I have no way of knowing.) Grow on plates with metal.
See what grows, how long does it take to lose resistance? Does it lose resistance at all?
After all that crap
Write the smegging lab report.
