Record the number of colonies on the plate.Carefully count the colonies on the plate having between 30 and 300 colonies.Following the incubation, inspect the pour plates and discern which of the plates contains between 30 and 300 colonies.Incubate Overnight at Optimal Temperature (You should check what the optimal temperature for growth is for your organism).Create Each Pour Plate Separately by Adding the Entire Contents of the Diluted Cell Culture to Warm TSA (Nutrient Agar) in a Petri Dish.Make sure to label your Petri dish with the dilution you used!.So, tube #6 has 1 / (10 x 10 x 10 x 10 x 10) dilution which is 1 / 10^5 or 1 / 100,000 or 1 : 100,000.Īfter you've made your serial dilutions, you will want to make some pour plates using a few of the most dilute concentrations from the series. Tube 6 has 1 / 10th of its volume that is composed of the 1 : 10,000 dilution from tube 5. I think you are beginning to see the pattern here! Then we take 1 ml from tube #5 and place it into tube #6.Tube 5 has 1 / 10th of its volume that is composed of the 1 : 1000 dilution from tube 4. You guessed it! We'll take 1 ml from tube #4 and place it into tube #5.Tube 4 has 1 / 10th of its volume that is composed of the 1 : 100 dilution from tube 3. Now, we can take 1 ml from tube #3 and place it into tube #4.Tube 3 has 1 / 10th of its volume that is composed of the 1 : 10 dilution from tube 2. Next, take 1 ml from tube #2 and place it into tube #3.Take 1 ml from the first tube and add it to the 9 ml of plain broth you have in tube 2.Then set up a number of tubes containing 9 ml each of broth alone (for dilutions).Start with 10 ml of your cell culture. This undiluted tube will be our tube #1.
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