Maheshri:Media
General Guidelines
To make 1L of liquid media:
1. In a 2L beaker dissolve everything EXCEPT the sugar (dextrose/galactose/raffinose), amino acids, nucleotides and antibiotics (if any) in ~500 mL water.
2. QS to the appropriate volume (Final volume - volume of sugar and amino acid solutions to be added) with water.
3. Autoclave for 40 min
4. Let cool in the 50°C water bath
5. Use sterile technique to add the sugar, amino acids, nucleotides and antibiotics. Mix.
To make 1L worth of agar plates (~ 40-50 yeast plates, 70-80 bacterial plates):
1. In a 2L beaker dissolve everything EXCEPT the sugar (dextrose/galactose/raffinose), amino acids, nucleotides and antibiotics (if any) in ~500 mL water.
2. QS to the appropriate volume (Final volume - volume of sugar and amino acid solutions to be added) with water.
3. Add 20g of Bacto agar (2% W/V)
4. Autoclave for 40 min
5. Let cool in the 50°C water bath
6. Use sterile technique to add the sugar, amino acids, nucleotides and antibiotics. Mix and pour.
Yeast Media
YePD/YPD
For 1L:
Peptone 20g
Yeast Extract 10g
50% Glucose 40mL
(Plate only) Agar 20g
YPAD,2X
For 1L:
Yeast Extract 20g
Peptone 40g
50% Glucose 80mL
Adenine 200mg
(Plate only) Agar 20g
SD min
For 1L:
Yeast Nitrogen Base w/o Amino Acids 6.7g
50% Glucose 40mL
(Plate only) Agar 20g
Amino Acid Solution, Complete
For 1L of 10x stock (good for 10L of media)
Adenine 0.4g Arginine 0.2g Histidine 0.2g Isoleucine 0.3g Leucine 1.0g Lysine 0.3g Methionine 0.2g Phenylalanine 0.5g Threonine 2.0g Tryptophan 0.4g Tyrosine 0.3g Uracil 0.2g Valine 1.5g
Bacterial Media
LB (Luria Broth)
For 1L:
Trypton 10g
Yeast Extract 5g
NaCl 10g
10N NaOH 100μL
(Plate only) Agar 20g
TB (Terrific Broth)
For 1L:
Trypton 10g
Yeast Extract 20g
Glycerol 4mL
(Plate only) Agar 20g
TB Amp plates
To make TB-Amp plates, add 1mL of 1000X ampicillin stock (at 100mg/mL) to 1L of TB media after it has been cooled down to 50°C.
