You will need to generate dropout plates to phenotype spore

You will need to generate dropout plates to phenotype spore colonies for each potential auxotrophy for the HOM3, TRP2, and ADE2 genes. Use the information in Table 2 to calculate the amount of supplements needed to make each dropout plate.

   Table 2. Supplement calculation chart.

Supplements

L-Threonine

L-Methionine

L-Tryptophan

Adenine

How to make 100 ml of stock solution

4g/100 ml

1g/100 ml

1g/100 ml

0.5g/100 ml

Stock concentration

(in mg/ml)

Stock concentration

(in x)

200x

500x

500x

200x

Working concentration

(in mg/ml)

Working concentration

(in x)

Volume to add to petri plate in µl (volume of petri plate = 25 ml)

Supplements	L-Threonine	L-Methionine	L-Tryptophan	Adenine
How to make 100 ml of stock solution	4g/100 ml	1g/100 ml	1g/100 ml	0.5g/100 ml
Stock concentration (in mg/ml)
Stock concentration (in x)	200x	500x	500x	200x
Working concentration (in mg/ml)
Working concentration (in x)
Volume to add to petri plate in µl (volume of petri plate = 25 ml)

Solution

Supplements

L-Threonine

L-Methionine

L-Tryptophan

Adenine

How to make 100 ml of stock solution

4g/100 ml

1g/100 ml

1g/100 ml

0.5g/100 ml

Stock concentration

(in mg/ml)

Stock concentration

(in x)

200x 40   

8000mg/ml

500x 10

5000mg/ml

500x 10

5000mg/ml

200x 5

1000mg/ml

Working concentration

(in mg/ml)

Working concentration

(in x)

Volume to add to petri plate in µl (volume of petri plate = 25 ml)

Calculation:

*Stock concentration (L-Threonine)

=4000 mg/100ml =40 mg/ml =1x concentration

*for 200x concentration = 200 x 40 = 8000mg/ml

*Stock concentration (L-Methionine)

=1000 mg/100ml =10 mg/ml =1x concentration

*for 500x concentration = 500 x 10 = 5000mg/ml

*Stock concentration (L-Tryptophan)

=1000 mg/100ml =10 mg/ml =1x concentration

*for 500x concentration = 500 x 10 = 5000mg/ml

*Stock concentration (Adenine)

=500 mg/100ml =5 mg/ml =1x concentration

*for 200x concentration = 200 x 5 = 1000mg/ml

Supplements	L-Threonine	L-Methionine	L-Tryptophan	Adenine
How to make 100 ml of stock solution	4g/100 ml	1g/100 ml	1g/100 ml	0.5g/100 ml
Stock concentration (in mg/ml)	40mg/ml	10mg/ml	10mg/ml	5mg/ml
Stock concentration (in x)	200x 40    8000mg/ml	500x 10 5000mg/ml	500x 10 5000mg/ml	200x 5 1000mg/ml
Working concentration (in mg/ml)	40 mg/ml	10mg/ml	10 mg/ml	5 mg/ml
Working concentration (in x)	0.5 ml	0.2 ml	0.2 ml	0.5 ml
Volume to add to petri plate in µl (volume of petri plate = 25 ml)	500 ul	50 ul	50 ul	500 ul