Paul.xu at 17:23, 25 November 2021

2021-11-25T17:23:08Z

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'''Commonly used bacterial E.coli growth media'''

 '''LB (Luria Bertani) Miller broth (1%NaCl)''': 1% peptone, 0.5% yeast extract, and 1% NaCl (0.17M)

for 1 L: 10 g Peptone, 5 g yeast extract, 10 g NaCl.

 '''LB (Luria Bertani) Lennox Broth (0.5% NaCl)''': 1% peptone, 0.5% yeast extract, and 0.5% NaCl

 '''SOB medium (Super Optimal Broth)''': 2% peptone, 0.5% Yeast extract, 10mM NaCl, 2.5mM KCl, 10mM MgCl2 , 10mM MgSO4 '''SOC medium '''(Super Optimal broth with Catabolic repressor): SOB + 20mM glucose

 '''2x YT broth''' (2x Yeast extract and Tryptone): 1.6% peptone, 1% yeast extract, and 0.5% NaCl

Per liter: 25 g LB powder, add 6 g of peptone, 5 g yeast extract.

'''20 L''': 500 g LB powder, 120 g peptone, 100 g yeast.

'''Make LB from 2X YT:  '''
31g 2XYT, + 15 g NaCl, + 4g Trypton, water to 2L.

310 g 2XYT, + 150 g NaCl, 40 g Trypton, water to 20 L

 '''TB (Terrific Broth) medium''': 1.2% peptone, 2.4% yeast extract, 72 mM K2HPO4, 17 mM KH2PO4 and 0.4% glycerol

12 g peptone, 24 g yeast extract, 4 ml glycerol,

'''TB Plus''': composition per liter: 12g Bacto Tryptone, 24g Yeast Extract, 0.4% (w/v) glycerol. Sterilization on autoclave followed by addition of: Na2HPO4 (25mM), KH2PO4 (25mM), NH4Cl (50mM), Na2SO4 (5mM), Glucose (0.05%), α-Lactose (0.2%), MgSO4.7H20 (2 mM).

 '''SB (Super Broth) medium''': 3.2% peptone, 2% yeast extract, and 0.5% NaCl. Optional: Before use, 10ml of sterile 1M MgCl2 and 10ml of sterile 1M MgSO4 are added.

 '''SOC I''' (Super Optimal Catobolite Repression), composition per liter: 20g Bacto Tryptone, 5g Yeast Extract, 2ml of 5M NaCl, 2.5ml of 1M KCl, 10ml of 1M MgCl2, 10ml of 1M MgSO4, 20ml of 1M glucose. Adjust to 1L with type I water and sterilize by autoclaving

'''Cold Spring:'''

Tryptone 2% (w/v) : 2 g

Yeast extract 0.5% (w/v) : 0.5 g

NaCl 10 mM: 250 ul of 4 M

KCl 2.5 mM: 2.5 ml of 1 M

MgCl2 10 mM: 1 ml of 1 M

Glucose 20 mM: 2 ml of 1 M  or 0.36g

Prepare a solution containing the first four reagents, sterilize at 121 °C, and then add sterile MgCl2 and glucose.

'''OD15'''--grow E coli to OD15

Per L ddH2O water, add

6g Na2HPO4.7H2O 3g KH2PO4 5g NaCl 1g NH4Cl 10g tryptone 10g peptone 40g yeast extract autoclave just before use, add 10ml 40% glycerol (-->0.4%, 4 g /1000 ml ) and 1 ml 1M MgSO4  (-->1 mM)

'''OD9 ''':  (for growing E coli to OD 9 after o/n growth at 37 oC using  shake flasks) K2HPO4 : 12.54 g/L

KH2PO4 : 2.31 g/L

Yeast extract: 12.0 g/L

NaCl: 5.0 g/L

MgSO4 : 0.24 g/L

Glycerol: <strike>D-Glucose: </strike>6.0 g/L

NH4Cl: 1.5 g/L

'''TB-Glycerol''':

Yeast extract: 24 g/L Peptone from casein:  12 g/L Glycerol: 5 g/L K2HPO4 12.54 g/L KH2PO4 2.31 g/L pH ad 7.0

Glycerol-based fed-batch fermentation medium containing for BL21 (DE3)

Glycerol: 20 g/L

Soya peptone: 5 g/L Thiamine: 0.3 g/L

KH2PO4: 13.5 g/L

(NH4)2HPO4: 4.0 g/L MgSO4·7H2O: 1.4 g/L

Citric acid: 1.7 g/L 10 ml/L of a trace metal solution:

FeSO4·7H2O: 10 g/L CaCl2: 2 g/L 

ZnSO4·7H2O: 2.2 g/L , MnSO4·4H2O: 0.5 g/L,

CuSO4·5H2O: 1.0 g/L ,

(NH4)6Mo24·4H2O: 0.1 g/L Na2B4O7·10H2O: 0.02 g/L  dissolved in 5 M HCl;

== '''High density bacterial E.coli growth media''' ==

 DNAGroTM medium: Support E.coli growth to OD600 = 30 - 50. Increase plasmid DNA yield 10 times over LB ProGroTM medium: Support E.coli growth to OD600 = 30 - 50. Increase protein yield 10 times over LB AutoXTM medium: Support E.coli growth to OD600 = 30 - 50. Increase protein yield 10 times over LB without adding IPTG DetoXTM medium: Reduce protein toxicity and support E.coli growth to OD600 = 30 InduXTM medium: Support E.coli growth to OD600 = 30. Increase protein yield, solubility, stability and activity SecProTM medium: Increase secretary protein expression over 100 times than LB or TB
[[Category:Buffer Recipe]]

Bacterial Medium - Revision history

Paul.xu at 17:23, 25 November 2021