SOB

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Adjust to pH 7.5 prior to use.  This requires approximately 25 ml of 1M NaOH per liter.
Adjust to pH 7.5 prior to use.  This requires approximately 25 ml of 1M NaOH per liter.
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==15/10 medium==
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Growth of competent TOP10 cells in Example 2 of the Bloom05 patent is performed in 15/10 broth, which is similar to SOB:
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* 1.5% yeast extract
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* 1% Bacto-Tryptone
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* 10mM NaCl
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* 2mM KCl
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* 10 mM MgCl<sub>2</sub>
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* 10 mM MgSO<sub>4</sub>
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==Extended instructions==
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*1 M MgCl2 stock: dissolve 20.33 g MgCl2 6H2O in 100 ml ddH2O (XXX), autoclave on liquid cycle @ XXX°C for 20 min (can be done at the same time as SOC pre-mix below)
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*250 mM KCl stock: dissolve 1.86 KCl in 100 ml ddH2O (XXX)
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*combine:
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{| border="1" cellpadding="5" cellspacing="0"
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|+
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! Reagent
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! for 1 L
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! 500 mL
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! 100 mL
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|-=
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| tryptone
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| 20 g
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| 10 g
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| 2g
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|-
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| yeast
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| 5g
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| 2.5 g
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|0.5 g
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|-
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| NaCl
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|0.5 g
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| 0.25 g
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|0.05 g
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|-
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| 250 nM KCl
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|10 mL
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|5mL
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|1 mL
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|-
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| ddH2O
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| 980 ml
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| 490 ml
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| 98 ml
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|}
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*adjust pH to 7.0 w/ NaOH
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*bring to volume:
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 +
{| border="1" cellpadding="5" cellspacing="0"
 +
|+
 +
! Reagent
 +
! for 1 L
 +
! 500 mL
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! 100 mL
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 +
|}
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*autoclave on liquid cycle @ XXX°C for 20 min
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*add autoclaved 1 M MgCl2
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 +
{| border="1" cellpadding="5" cellspacing="0"
 +
|+
 +
! Reagent
 +
! for 1 L
 +
! 500 mL
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! 100 mL
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|-=
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| 1 M MgCl2
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| 10 ml
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| 5 ml
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| 1 ml
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|}
==Source==
==Source==
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==Trivia==
==Trivia==
SOB supposedly stands for super optimal broth.
SOB supposedly stands for super optimal broth.
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 +
[[Category:Material]]  [[Category:Standard Media]]

Current revision

Contents

Summary

SOB Medium. Used in growing bacteria for preparing chemically competent cells

Ingredients

  • 0.5% (w/v) yeast extract
  • 2% (w/v) tryptone
  • 10 mM NaCl
  • 2.5 mM KCl
  • 20 mM MgSO4

Per liter:

  • 5 g yeast extract
  • 20 g tryptone
  • 0.584 g NaCl
  • 0.186 g KCl
  • 2.4 g MgSO4

Note: Some formulations of SOB use 10 mM MgCl2 and 10 mM MgSO4 instead of 20 mM MgSO4.

SOB medium is also available dry premixed from Difco, 0443-17.

Adjust to pH 7.5 prior to use. This requires approximately 25 ml of 1M NaOH per liter.

15/10 medium

Growth of competent TOP10 cells in Example 2 of the Bloom05 patent is performed in 15/10 broth, which is similar to SOB:

  • 1.5% yeast extract
  • 1% Bacto-Tryptone
  • 10mM NaCl
  • 2mM KCl
  • 10 mM MgCl2
  • 10 mM MgSO4

Extended instructions

  • 1 M MgCl2 stock: dissolve 20.33 g MgCl2 6H2O in 100 ml ddH2O (XXX), autoclave on liquid cycle @ XXX°C for 20 min (can be done at the same time as SOC pre-mix below)
  • 250 mM KCl stock: dissolve 1.86 KCl in 100 ml ddH2O (XXX)
  • combine:
Reagent for 1 L 500 mL 100 mL
tryptone 20 g 10 g 2g
yeast 5g 2.5 g 0.5 g
NaCl 0.5 g 0.25 g 0.05 g
250 nM KCl 10 mL 5mL 1 mL
ddH2O 980 ml 490 ml 98 ml
  • adjust pH to 7.0 w/ NaOH
  • bring to volume:
Reagent for 1 L 500 mL 100 mL
  • autoclave on liquid cycle @ XXX°C for 20 min
  • add autoclaved 1 M MgCl2
Reagent for 1 L 500 mL 100 mL
1 M MgCl2 10 ml 5 ml 1 ml

Source

Adapted From:

F. Ausubel et al., Short Protocols in Molecular Biology (John Wiley & Sons, ed. 4, 1999) pg. A1-36

Trivia

SOB supposedly stands for super optimal broth.

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