SYMINGTON LAB Media Recipes

How to make YPD (YAPD) Liquid:

 For 2L solution:

Add:                                                                            

1750mL dH20 from Milli-Q

20g Yeast Extract

40g Bacto Peptone

40g Glucose

10mL of 0.2% Adenine

Then:   

Stir using hot plate w/ stir bar for ~30min.

 Next, add MQ water up to 2000mL.

 Pour into 1L glass bottles. Each container should have ~650mL media.

Autoclave: ~40min

How to make YP Liquid:

For 4L solution:

Add:                                                                            

3000mL dH20 from Milli-Q

40g Yeast Extract

80g Bacto Peptone

 Then:       

Stir using hot plate w/ stir bar for ~30min.

Next, add MQ water up to 3600mL.

 Pour into 1L glass bottles. Each container should have ~900mL media.

 Autoclave: ~40min  

How to make LB Liquid:

 For 2L solution:

Add:                                                                                                     

1750mL dH20 from Milli-Q

10g Yeast Extract

20g Tryptone

10g NaCl

 Then:    

Stir using hot plate w/ stir bar for ~30min.

Next, add MQ water up to 2000mL.

Pour into 1L glass bottles. Each container should have ~500mL media.

Autoclave: ~40min

How to make YPD (YAPD) Plates:

For (0.5L) or 1L

Add:                                                                                                     

(0.5L) 1L dH20 from Milli-Q

(5g) 10g Yeast Extract

(10g) 20g Bacto Peptone

(10g) 20g Glucose

(10g) 20g Bacto Agar

(2.5mL) 5mL0.2% Adenine

 Autoclave: ~35-45min

 Then:

Cool on hot plate with spinner ~20-30min.

Pour plates. Let dry at room temperature for 2 days.

How to make Dissection Plates:

 For (0.5L) or 1L

Add:                                                                                                     

(0.5L) 1L dH20 from Milli-Q

(5g) 10g Yeast Extract

(10g) 20g Bacto Peptone

(10g) 20g Glucose

(10g) 20g NOBLE Agar

(2.5mL) 5mL 0.2% Adenine

 Autoclave: ~30min

 Then:

Cool on hot plate with spinner ~20-30min.

Pipette 23mL into each plates (before the agar settles!)

Pour plates. Let dry at room temperature for 3 days.

How to make YP-Galactose Plates:

 For (0.5L) or 1L

Add:                                                                                                     

(0.45L) 900mL dH20 from Milli-Q

 (5g) 10g Yeast Extract

(10g) 20g Bacto Peptone

(10g) 20g Bacto Agar

Autoclave: ~30min

 Cool on hot plate with spinner ~20-30min.

 Now add:

(50mL) 100mL 20% Galactose

Stir >5min to allow solution to mix well.

Pour plates. Let dry at room temperature for 2 days.

How to make SC-Trp Plates:

For (0.5L) or 1L

Add:                                                                                                     

(0.5L) 1L dH20 from Milli-Q

(0.96g) 1.92g SC-Trp drop-out powder

(3.35g) 6.7g YNB

(10g) 20g Glucose

(10g) 20g Bacto Agar

 Autoclave: ~30min 

 Cool on hot plate with spinner ~20-30min.

Pour plates. Let dry at room temperature for 2 days.

How to make LB+AMP Plates:

For (0.5L) or 1L

Add:                                                                                                     

(2.5g) 5g Yeast Extract

(5g) 10g Tryptone

(5g) 10g NaCl

(7.5g) 15g Bacto Agar

 Autoclave: ~40min 

Cool on hot plate with spinner ~20-30min.

 Now Add:

(0.5mL) 1mL 100mg/mL Ampicillin (Thawed) (we make -20C freezer aliquots)

 Pour plates. Let dry at room temperature for 2-3 days.

How to make YPD+ G418 Plates:

For (0.5L) or 1L

Add:                                                                                                     

(0.5L) 1L dH20 from Milli-Q

(5g) 10g Yeast Extract

(10g) 20g Bacto Peptone

(10g) 20g Glucose

(10g) 20g Bacto Agar

(2.5mL) 5mL 0.2% Adenine

 Autoclave: ~30 min

 Then:

Cool on hot plate with spinner ~20-30min.

Now add:

(2mL) 4mL 50mg/mL G418 solution

 Stir >5min to allow solution to mix well.

Pour plates. Let dry at room temperature for 2 days.

How to make YPD+ HYG Plates:

For (0.5L) or 1L

Add:                                                                                                     

(0.5L) 1L dH20 from Milli-Q

(5g) 10g Yeast Extract

(10g) 20g Bacto Peptone

(10g) 20g Glucose

(10g) 20g Bacto Agar

(2.5mL) 5mL 0.2% Adenine

 Autoclave: ~30 min

 Then:

Cool on hot plate with spinner ~20-30min.

Now add:

(300uL) 600uL Hygromycin B

Stir >5min to allow solution to mix well.

Pour plates. Let dry at room temperature for 2 days.

How to make YPD+ NAT Plates:

For (0.5L) or 1L

Add:                                                                                                     

(0.5L) 1L dH20 from Milli-Q

(5g) 10g Yeast Extract

(10g) 20g Bacto Peptone

(10g) 20g Glucose

(10g) 20g Bacto Agar

(2.5mL) 5mL 0.2% Adenine

 Autoclave: ~30 min

 Then:

Cool on hot plate with spinner ~20-30min.

Now add:

(250uL) 500uL Clon-NAT (Thawed) (-20C freezer aliquots)

Stir >5min to allow solution to mix well.

Pour plates. Let dry at room temperature for 2 days.