Monday June 11th: All in A Day's Work
On Monday, I plated three samples of each dilution for the four different brands of Kefir.
I plated more Kefir so that we could count the number of colonies in a single sample.
The previous plates were used as reference to see what dilution factor we would need
In addition to plating, I had also count the samples taken from La Yogurt at dilution factor 105.
The average amount of colonies for this particular brand was 62.7 x 106 CFU/ mL.
To get a better understanding of how we count colonies, please refer to the post
titled “How to Count Bacterial Colonies”.
I plated more Kefir so that we could count the number of colonies in a single sample.
The previous plates were used as reference to see what dilution factor we would need
to plate to be able to count the colonies.
The average amount of colonies for this particular brand was 62.7 x 106 CFU/ mL.
To get a better understanding of how we count colonies, please refer to the post
titled “How to Count Bacterial Colonies”.
Now the fun stuff!! Upon taking a look at the plates that had been in the incubator over the weekend, Professor Murthi and I decided different dilutions for different brands.
Latta: dilute to 103 and plate the 102 and 103
Fresh Made: dilute to 104 and plate the 103 and 104
La Yogurt: dilute to 105 and plate 105
Lifeway: dilute to 2x and 5x
Diluting Latta Method:
1. Label the test tubes by their dilution factors (10, 102, 103)
2. Using the micropipette, add 1800 μL of LB broth to each of the three test tubes .
3. Add 200 μL of Latta kefir to test tube 1 (dilution factor 10). Shake the solution.
4. From test tube 1 (dilution factor of 10), extract 200 μL and carefully add it to the next test tube (test tube 2/ dilution factor of 102). Shake test tube 2.
5. From test tube 2 (dilution factor of 102), extract 200 μL and carefully add it to the next test tube (test tube 3/ dilution factor of 103). Shake test tube 3.
6. Extract 100 μL of 102 and place it in the middle of plate labeled “Latta 102 6/11”. Use the L-shaped spreader until all of the liquid is absorbed.
7. Repeat this process two more times so that there is a total of 3 plates for the 102 dilution factor for Latta.
8. Extract 100 μL of 103 and place it in the middle of plate labeled “Latta 103 6/11”. Use the L-shaped spreader until all of the liquid is absorbed.
9. Repeat this process two more times so that there is a total of 3 plates for the 103 dilution factor for Latta.
1. Label the test tubes by their dilution factors (10, 102, 103)
2. Using the micropipette, add 1800 μL of LB broth to each of the three test tubes .
3. Add 200 μL of Latta kefir to test tube 1 (dilution factor 10). Shake the solution.
4. From test tube 1 (dilution factor of 10), extract 200 μL and carefully add it to the next test tube (test tube 2/ dilution factor of 102). Shake test tube 2.
5. From test tube 2 (dilution factor of 102), extract 200 μL and carefully add it to the next test tube (test tube 3/ dilution factor of 103). Shake test tube 3.
6. Extract 100 μL of 102 and place it in the middle of plate labeled “Latta 102 6/11”. Use the L-shaped spreader until all of the liquid is absorbed.
7. Repeat this process two more times so that there is a total of 3 plates for the 102 dilution factor for Latta.
8. Extract 100 μL of 103 and place it in the middle of plate labeled “Latta 103 6/11”. Use the L-shaped spreader until all of the liquid is absorbed.
9. Repeat this process two more times so that there is a total of 3 plates for the 103 dilution factor for Latta.
Diluting Fresh Made Method:
1. Label the test tubes by their dilution factors (10, 102, 103, 104)
2. Using the micropipette, add 1800 μL of LB broth to each of the four test tubes .
3. Add 200 μL of Fresh Made kefir to test tube 1 (dilution factor 10). Shake the solution.
4. From test tube 1 (dilution factor of 10), extract 200 μL and carefully add it to the next test tube (test tube 2/ dilution factor of 102). Shake test tube 2.
5. From test tube 2 (dilution factor of 102), extract 200 μL and carefully add it to the next test tube (test tube 3/ dilution factor of 103). Shake test tube 3.
6. From test tube 3 (dilution factor of 103), extract 200 μL and carefully add it to the next test tube (test tube 4/ dilution factor of 104). Shake test tube 4.
7. Extract 100 μL of 103 and place it in the middle of plate labeled “Fresh Made 103 6/11”. Use the L-shaped spreader until all of the liquid is absorbed.
8. Repeat this process two more times so that there is a total of 3 plates for the 103 dilution factor for Fresh Made.
9. Extract 100 μL of 104 and place it in the middle of plate labeled “Fresh Made 104 6/11”. Use the L-shaped spreader until all of the liquid is absorbed.
10. Repeat this process two more times so that there is a total of 3 plates for the 104 dilution factor for Fresh Made.
1. Label the test tubes by their dilution factors (10, 102, 103, 104)
2. Using the micropipette, add 1800 μL of LB broth to each of the four test tubes .
3. Add 200 μL of Fresh Made kefir to test tube 1 (dilution factor 10). Shake the solution.
4. From test tube 1 (dilution factor of 10), extract 200 μL and carefully add it to the next test tube (test tube 2/ dilution factor of 102). Shake test tube 2.
5. From test tube 2 (dilution factor of 102), extract 200 μL and carefully add it to the next test tube (test tube 3/ dilution factor of 103). Shake test tube 3.
6. From test tube 3 (dilution factor of 103), extract 200 μL and carefully add it to the next test tube (test tube 4/ dilution factor of 104). Shake test tube 4.
7. Extract 100 μL of 103 and place it in the middle of plate labeled “Fresh Made 103 6/11”. Use the L-shaped spreader until all of the liquid is absorbed.
8. Repeat this process two more times so that there is a total of 3 plates for the 103 dilution factor for Fresh Made.
9. Extract 100 μL of 104 and place it in the middle of plate labeled “Fresh Made 104 6/11”. Use the L-shaped spreader until all of the liquid is absorbed.
10. Repeat this process two more times so that there is a total of 3 plates for the 104 dilution factor for Fresh Made.
Diluting La Yogurt Method:
1. Label the test tubes by their dilution factors (10, 102, 103, 104, 105)
2. Using the micropipette, add 1800 μL of LB broth to each of the four test tubes .
3. Add 200 μL of La Yogurt kefir to test tube 1 (dilution factor 10). Shake the solution.
4. From test tube 1 (dilution factor of 10), extract 200 μL and carefully add it to the next test tube (test tube 2/ dilution factor of 102). Shake test tube 2.
5. From test tube 2 (dilution factor of 102), extract 200 μL and carefully add it to the next test tube (test tube 3/ dilution factor of 103). Shake test tube 3.
6. From test tube 3 (dilution factor of 103), extract 200 μL and carefully add it to the next test tube (test tube 4/ dilution factor of 104). Shake test tube 4.
7. From test tube 3 (dilution factor of 103), extract 200 μL and carefully add it to the next test tube (test tube 4/ dilution factor of 104). Shake test tube 4.
8. From test tube 4 (dilution factor of 104), extract 200 μL and carefully add it to the next test tube (test tube 5/ dilution factor of 105). Shake test tube 5.
9. Extract 100 μL of 105 and place it in the middle of plate labeled “La Yogurt 105 6/11”. Use the L-shaped spreader until all of the liquid is absorbed.
10. Repeat this process two more times so that there is a total of 3 plates for the 105 dilution factor for La Yogurt.
1. Label the test tubes by their dilution factors (10, 102, 103, 104, 105)
2. Using the micropipette, add 1800 μL of LB broth to each of the four test tubes .
3. Add 200 μL of La Yogurt kefir to test tube 1 (dilution factor 10). Shake the solution.
4. From test tube 1 (dilution factor of 10), extract 200 μL and carefully add it to the next test tube (test tube 2/ dilution factor of 102). Shake test tube 2.
5. From test tube 2 (dilution factor of 102), extract 200 μL and carefully add it to the next test tube (test tube 3/ dilution factor of 103). Shake test tube 3.
6. From test tube 3 (dilution factor of 103), extract 200 μL and carefully add it to the next test tube (test tube 4/ dilution factor of 104). Shake test tube 4.
7. From test tube 3 (dilution factor of 103), extract 200 μL and carefully add it to the next test tube (test tube 4/ dilution factor of 104). Shake test tube 4.
8. From test tube 4 (dilution factor of 104), extract 200 μL and carefully add it to the next test tube (test tube 5/ dilution factor of 105). Shake test tube 5.
9. Extract 100 μL of 105 and place it in the middle of plate labeled “La Yogurt 105 6/11”. Use the L-shaped spreader until all of the liquid is absorbed.
10. Repeat this process two more times so that there is a total of 3 plates for the 105 dilution factor for La Yogurt.
Diluting Lifeway Method:
1. Obtain two test tubes and label them by their dilution factor. These tubes tubes will either be a 2x or a 5x dilution.
2. In the 2x dilution test tube, add 1.0 mL of LB.
3. Add 1.0 mL of Lifeway Kefir to the 2x dilution test tube. Shake.
4. Extract 100 μL of the 2x dilution into the plate labeled “Lifeway 2x 6/11”. Use the L-shaped spreaders until all of the liquid has been absorbed.
5. Repeat this process two more times so that there is a total of 3 plates for the 2x dilution factor for Lifeway.
6. In the 5x dilution test tube, add 4.0 ml of LB.
7. Add 1.0 mL of Lifeway Kefir to the 5x dilution test tube. Shake.
8. Extract 100 μL of the 5x dilution into the plate labeled “Lifeway 5x 6/11”. Use the L-shaped spreaders until all of the liquid has been absorbed.
9. Repeat this process two more times so that there is a total of 3 plates for the 5x dilution factor for Lifeway.
1. Obtain two test tubes and label them by their dilution factor. These tubes tubes will either be a 2x or a 5x dilution.
2. In the 2x dilution test tube, add 1.0 mL of LB.
3. Add 1.0 mL of Lifeway Kefir to the 2x dilution test tube. Shake.
4. Extract 100 μL of the 2x dilution into the plate labeled “Lifeway 2x 6/11”. Use the L-shaped spreaders until all of the liquid has been absorbed.
5. Repeat this process two more times so that there is a total of 3 plates for the 2x dilution factor for Lifeway.
6. In the 5x dilution test tube, add 4.0 ml of LB.
7. Add 1.0 mL of Lifeway Kefir to the 5x dilution test tube. Shake.
8. Extract 100 μL of the 5x dilution into the plate labeled “Lifeway 5x 6/11”. Use the L-shaped spreaders until all of the liquid has been absorbed.
9. Repeat this process two more times so that there is a total of 3 plates for the 5x dilution factor for Lifeway.
All of the plates were placed in the incubator at 37 degrees Celsius where they will grow for 48 hours. Hopefully we will see a lot of growth after they are taken out!
Comments
Post a Comment