Maheshri:Media

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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:

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

YPAD,2X

For 1L:

Yeast Extract 20g

Peptone 40g

50% Glucose 80mL

Adenine 200mg

SD min

For 1L:

Yeast Nitrogen Base w/o Amino Acids 6.7g

50% Glucose 40mL

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

TB (Terrific Broth)

For 1L:

Trypton 10g

Yeast Extract 20g

Glycerol 4mL