M10 Medium - Revision history

Madachi: /* Method */

2026-10-30T17:11:08Z

Method

← Older revision		Revision as of 17:11, 30 October 2026
Line 48:		Line 48:

	<sup>c</sup> The volatile fatty acid mixture used was the following: acetic, 17 ml (2.9 X 10<sup>-2</sup> M); propionic, 6ml (8.0 X 10<sup>-</sup><sup>3</sup> M); butyric, 4 ml (4.3 X 10<sup>-</sup><sup>3</sup> M); isobutyric, 1 ml (1.1 X 10<sup>-3</sup> M); n-valeric, isovaleric, and DL-α-methylbutyric, 1 ml each (9 X 10<sup>-4</sup> M). The figures in parentheses indicate the final concentration of each acid when 0.31 ml of mixture was placed in a final medium volume of 100 ml.		<sup>c</sup> The volatile fatty acid mixture used was the following: acetic, 17 ml (2.9 X 10<sup>-2</sup> M); propionic, 6ml (8.0 X 10<sup>-</sup><sup>3</sup> M); butyric, 4 ml (4.3 X 10<sup>-</sup><sup>3</sup> M); isobutyric, 1 ml (1.1 X 10<sup>-3</sup> M); n-valeric, isovaleric, and DL-α-methylbutyric, 1 ml each (9 X 10<sup>-4</sup> M). The figures in parentheses indicate the final concentration of each acid when 0.31 ml of mixture was placed in a final medium volume of 100 ml.
	== ~~Method~~ ==		== Direction ==
	Mix all the components in distilled water except Na<sub>2</sub>CO<sub>3</sub> and Cystein·HCl·H<sub>2</sub>O. Boil the medium, then cool it immediately in an ice bath while bubuling with 100% CO<sub>2</sub>. Add appropirate amount of Na<sub>2</sub>CO<sub>3</sub> and Cystein·HCl·H<sub>2</sub>O, once the medium is cooled. Adjust th pH to 6.7-6.8 by addition of NaOH. Aliquot the medium, and autoclave at 121 °C for 20 minutes.		Mix all the components in distilled water except Na<sub>2</sub>CO<sub>3</sub> and Cystein·HCl·H<sub>2</sub>O. Boil the medium, then cool it immediately in an ice bath while bubuling with 100% CO<sub>2</sub>. Add appropirate amount of Na<sub>2</sub>CO<sub>3</sub> and Cystein·HCl·H<sub>2</sub>O, once the medium is cooled. Adjust th pH to 6.7-6.8 by addition of NaOH. Aliquot the medium, and autoclave at 121 °C for 20 minutes.

Madachi: /* Method */

2026-10-29T22:18:07Z

Method

← Older revision		Revision as of 22:18, 29 October 2026
Line 48:		Line 48:

	<sup>c</sup> The volatile fatty acid mixture used was the following: acetic, 17 ml (2.9 X 10<sup>-2</sup> M); propionic, 6ml (8.0 X 10<sup>-</sup><sup>3</sup> M); butyric, 4 ml (4.3 X 10<sup>-</sup><sup>3</sup> M); isobutyric, 1 ml (1.1 X 10<sup>-3</sup> M); n-valeric, isovaleric, and DL-α-methylbutyric, 1 ml each (9 X 10<sup>-4</sup> M). The figures in parentheses indicate the final concentration of each acid when 0.31 ml of mixture was placed in a final medium volume of 100 ml.		<sup>c</sup> The volatile fatty acid mixture used was the following: acetic, 17 ml (2.9 X 10<sup>-2</sup> M); propionic, 6ml (8.0 X 10<sup>-</sup><sup>3</sup> M); butyric, 4 ml (4.3 X 10<sup>-</sup><sup>3</sup> M); isobutyric, 1 ml (1.1 X 10<sup>-3</sup> M); n-valeric, isovaleric, and DL-α-methylbutyric, 1 ml each (9 X 10<sup>-4</sup> M). The figures in parentheses indicate the final concentration of each acid when 0.31 ml of mixture was placed in a final medium volume of 100 ml.
	=== Method ===		== Method ==
	Mix all the components in distilled water except Na<sub>2</sub>CO<sub>3</sub> and Cystein·HCl·H<sub>2</sub>O. Boil the medium, then cool it immediately in an ice bath while bubuling with 100% CO<sub>2</sub>. Add appropirate amount of Na<sub>2</sub>CO<sub>3</sub> and Cystein·HCl·H<sub>2</sub>O, once the medium is cooled. Adjust th pH to 6.7-6.8 by addition of NaOH. Aliquot the medium, and autoclave at 121 °C for 20 minutes.		Mix all the components in distilled water except Na<sub>2</sub>CO<sub>3</sub> and Cystein·HCl·H<sub>2</sub>O. Boil the medium, then cool it immediately in an ice bath while bubuling with 100% CO<sub>2</sub>. Add appropirate amount of Na<sub>2</sub>CO<sub>3</sub> and Cystein·HCl·H<sub>2</sub>O, once the medium is cooled. Adjust th pH to 6.7-6.8 by addition of NaOH. Aliquot the medium, and autoclave at 121 °C for 20 minutes.

Madachi: /* Sources */

2026-10-29T22:09:16Z

Sources

← Older revision		Revision as of 22:09, 29 October 2026
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	== Sources ==		== Sources ==

	[1] ~~Hobson PN~~. ~~(1969)~~. ~~Rumen~~ bacteria. In: ~~''Methods in Microbiology'', vol 3B. Academic Press, pp. 133–149~~.		[1] Caldwell DR, Bryant MP. 1966. Medium without rumen fluid for nonselective enumeration and isolation of rumen bacteria. Appl Microbiol 14:794–801.

Madachi at 22:05, 29 October 2026

2026-10-29T22:05:18Z

← Older revision		Revision as of 22:05, 29 October 2026
Line 48:		Line 48:

	<sup>c</sup> The volatile fatty acid mixture used was the following: acetic, 17 ml (2.9 X 10<sup>-2</sup> M); propionic, 6ml (8.0 X 10<sup>-</sup><sup>3</sup> M); butyric, 4 ml (4.3 X 10<sup>-</sup><sup>3</sup> M); isobutyric, 1 ml (1.1 X 10<sup>-3</sup> M); n-valeric, isovaleric, and DL-α-methylbutyric, 1 ml each (9 X 10<sup>-4</sup> M). The figures in parentheses indicate the final concentration of each acid when 0.31 ml of mixture was placed in a final medium volume of 100 ml.		<sup>c</sup> The volatile fatty acid mixture used was the following: acetic, 17 ml (2.9 X 10<sup>-2</sup> M); propionic, 6ml (8.0 X 10<sup>-</sup><sup>3</sup> M); butyric, 4 ml (4.3 X 10<sup>-</sup><sup>3</sup> M); isobutyric, 1 ml (1.1 X 10<sup>-3</sup> M); n-valeric, isovaleric, and DL-α-methylbutyric, 1 ml each (9 X 10<sup>-4</sup> M). The figures in parentheses indicate the final concentration of each acid when 0.31 ml of mixture was placed in a final medium volume of 100 ml.
	=== ~~Directions~~ ===		=== Method ===
	Mix all the components in distilled water except Na<sub>2</sub>CO<sub>3</sub> and Cystein·HCl·H<sub>2</sub>O. Boil the medium, then cool it immediately in an ice bath while bubuling with 100% CO<sub>2</sub>. Add appropirate amount of Na<sub>2</sub>CO<sub>3</sub> and Cystein·HCl·H<sub>2</sub>O, once the medium is cooled. Adjust th pH to 6.7-6.8 by addition of NaOH. Aliquot the medium, and autoclave at 121 °C for 20 minutes.		Mix all the components in distilled water except Na<sub>2</sub>CO<sub>3</sub> and Cystein·HCl·H<sub>2</sub>O. Boil the medium, then cool it immediately in an ice bath while bubuling with 100% CO<sub>2</sub>. Add appropirate amount of Na<sub>2</sub>CO<sub>3</sub> and Cystein·HCl·H<sub>2</sub>O, once the medium is cooled. Adjust th pH to 6.7-6.8 by addition of NaOH. Aliquot the medium, and autoclave at 121 °C for 20 minutes.

Line 55:		Line 55:
	== Sources ==		== Sources ==

	[1] ~~Caldwell DR, Bryant MP~~. ~~1966~~. ~~Medium without rumen fluid for nonselective enumeration and isolation of rumen~~ bacteria. ~~Appl Microbiol 14~~:~~794–801~~.		[1] Hobson PN. (1969). Rumen bacteria. In: ''Methods in Microbiology'', vol 3B. Academic Press, pp. 133–149.

Madachi: /* Directions */

2026-10-22T22:11:35Z

Directions

← Older revision		Revision as of 22:11, 22 October 2026
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	<sup>a</sup> Weight/ volume unless otherwise indicated.		<sup>a</sup> Weight/ volume unless otherwise indicated.

	<sup>b</sup> Except for K<sub>2</sub>HPO<sub>4</sub>, which was prepared separately, the minerals were prepared as a common stock solution. The final mineral concentrations in the medium were as follows: KH<sub>2</sub>PO<sub>4</sub>, 1.3 X 10<sup>-3</sup> M; NaCl, 7.6 X 10<sup>-4</sup> M; (NH)<sub>2</sub>SO<sub>4</sub>, 3.4 X 10<sup>-3</sup> M; K<sub>2</sub>HPO<sub>4</sub>, 1.7 X 10<sup>-3</sup> M; Ca(Cl)<sub>2</sub>, 4.1 X 10<sup>-4</sup> M; MgSO<sub>4</sub>~~c·7H~~<sub>2</sub>0, 3.8 X 10<sup>-4</sup> M.		<sup>b</sup> Except for K<sub>2</sub>HPO<sub>4</sub>, which was prepared separately, the minerals were prepared as a common stock solution. The final mineral concentrations in the medium were as follows: KH<sub>2</sub>PO<sub>4</sub>, 1.3 X 10<sup>-3</sup> M; NaCl, 7.6 X 10<sup>-4</sup> M; (NH)<sub>2</sub>SO<sub>4</sub>, 3.4 X 10<sup>-3</sup> M; K<sub>2</sub>HPO<sub>4</sub>, 1.7 X 10<sup>-3</sup> M; Ca(Cl)<sub>2</sub>, 4.1 X 10<sup>-4</sup> M; MgSO<sub>4</sub>·7H<sub>2</sub>0, 3.8 X 10<sup>-4</sup> M.

	<sup>c</sup> The volatile fatty acid mixture used was the following: acetic, 17 ml (2.9 X 10<sup>-2</sup> M); propionic, 6ml (8.0 X 10<sup>-</sup><sup>3</sup> M); butyric, 4 ml (4.3 X 10<sup>-</sup><sup>3</sup> M); isobutyric, 1 ml (1.1 X 10<sup>-3</sup> M); n-valeric, isovaleric, and DL-α-methylbutyric, 1 ml each (9 X 10<sup>-4</sup> M). The figures in parentheses indicate the final concentration of each acid when 0.31 ml of mixture was placed in a final medium volume of 100 ml.		<sup>c</sup> The volatile fatty acid mixture used was the following: acetic, 17 ml (2.9 X 10<sup>-2</sup> M); propionic, 6ml (8.0 X 10<sup>-</sup><sup>3</sup> M); butyric, 4 ml (4.3 X 10<sup>-</sup><sup>3</sup> M); isobutyric, 1 ml (1.1 X 10<sup>-3</sup> M); n-valeric, isovaleric, and DL-α-methylbutyric, 1 ml each (9 X 10<sup>-4</sup> M). The figures in parentheses indicate the final concentration of each acid when 0.31 ml of mixture was placed in a final medium volume of 100 ml.
	=== Directions ===		=== Directions ===
	~~Mixe~~ all the components in distilled water except Na<sub>2</sub>CO<sub>3</sub> and Cystein·HCl·H<sub>2</sub>O. Boil the medium, then cool it immediately in an ice bath ~~under bubbled~~ with 100% CO<sub>2</sub>. Add appropirate amount of Na<sub>2</sub>CO<sub>3</sub> and Cystein·HCl·H<sub>2</sub>O, once the medium is cooled. Adjust th pH to 6.7 ~ 6.8 by addition of NaOH. Aliquot the medium, and autoclave at 121 °C for 20 minutes.		Mix all the components in distilled water except Na<sub>2</sub>CO<sub>3</sub> and Cystein·HCl·H<sub>2</sub>O. Boil the medium, then cool it immediately in an ice bath while bubuling with 100% CO<sub>2</sub>. Add appropirate amount of Na<sub>2</sub>CO<sub>3</sub> and Cystein·HCl·H<sub>2</sub>O, once the medium is cooled. Adjust th pH to 6.7-6.8 by addition of NaOH. Aliquot the medium, and autoclave at 121 °C for 20 minutes.

	== Notes ==		== Notes ==

Madachi at 22:05, 22 October 2026

2026-10-22T22:05:06Z

← Older revision		Revision as of 22:05, 22 October 2026
Line 48:		Line 48:

	<sup>c</sup> The volatile fatty acid mixture used was the following: acetic, 17 ml (2.9 X 10<sup>-2</sup> M); propionic, 6ml (8.0 X 10<sup>-</sup><sup>3</sup> M); butyric, 4 ml (4.3 X 10<sup>-</sup><sup>3</sup> M); isobutyric, 1 ml (1.1 X 10<sup>-3</sup> M); n-valeric, isovaleric, and DL-α-methylbutyric, 1 ml each (9 X 10<sup>-4</sup> M). The figures in parentheses indicate the final concentration of each acid when 0.31 ml of mixture was placed in a final medium volume of 100 ml.		<sup>c</sup> The volatile fatty acid mixture used was the following: acetic, 17 ml (2.9 X 10<sup>-2</sup> M); propionic, 6ml (8.0 X 10<sup>-</sup><sup>3</sup> M); butyric, 4 ml (4.3 X 10<sup>-</sup><sup>3</sup> M); isobutyric, 1 ml (1.1 X 10<sup>-3</sup> M); n-valeric, isovaleric, and DL-α-methylbutyric, 1 ml each (9 X 10<sup>-4</sup> M). The figures in parentheses indicate the final concentration of each acid when 0.31 ml of mixture was placed in a final medium volume of 100 ml.


	~~The final pH of media is 6.7 to 6.8.~~

	=== Directions ===		=== Directions ===
	~~Suspend 38 g of RCM~~ in ~~1 liter of~~ distilled water. Boil to ~~dissolve solids completely, aliquot~~, and ~~sterilize by autoclaving~~ at 121 °C for 15 minutes.		Mixe all the components in distilled water except Na<sub>2</sub>CO<sub>3</sub> and Cystein·HCl·H<sub>2</sub>O. Boil the medium, then cool it immediately in an ice bath under bubbled with 100% CO<sub>2</sub>. Add appropirate amount of Na<sub>2</sub>CO<sub>3</sub> and Cystein·HCl·H<sub>2</sub>O, once the medium is cooled. Adjust th pH to 6.7 ~ 6.8 by addition of NaOH. Aliquot the medium, and autoclave at 121 °C for 20 minutes.

	== Notes ==		== Notes ==
	[[File:Cell pellet formation of Megasphaera elsdenii strain AG22635 in RCM variant.png\|thumb\|Cell pellet of Megasphaera elsdenii strain AG22635 after a 10-minute 5000 x g centrifuge spin. The culture on the left was grown in commercial RCM. The culture on the right was grown in lab-made RCM without agar.]]
	Store bought RCM contains 0.5 g/L agar to mitigate oxygen permeability into the medium. The addition of this agar can have a compounding effect on the formation of [[wikipedia:Extracellular_polymeric_substance\|extracellular polymeric substances (EPS)]] in some species like [[Sharpea azabuensis KH1P5\|Sharpea azubuensis]] and [[Megasphaera elsdenii T81\|Megasphaera elsdeni]]. This EPS generation makes the formation of cell pellets difficult and can affect the procedure of making competent cells. It's recommended to make your RCM "in house" without agar to avoid this complication.










	~~<gallery>~~
	~~</gallery>~~

	== Sources ==		== Sources ==

	* [~~https://documents~~.~~thermofisher~~.~~com/TFS-Assets/MBD/Specification-Sheets/BT-SPEC-0099~~.~~pdf ThermoFisher CM0149] (Used by Guss Lab)~~		[1] Caldwell DR, Bryant MP. 1966. Medium without rumen fluid for nonselective enumeration and isolation of rumen bacteria. Appl Microbiol 14:794–801.

Madachi: Created page with "== Description == M10 is a medium without rumen fluid for nonselective enumeration and isolation of rumen bacteria [1]. == Recipe == {| class="wikitable" |+ !Component !Percentage in medium^a |- |Glucose |0.05 |- |Cellobiose |0.05 |- |Soluble Starch |0.05 |- |Minerals (v/v)^b |3.75 |- |Na₂S·9H₂O |0.025 |- |Cystein·HCl·H₂O |0.025 |- |Resazurin |0.001 |- |Na₂CO₃ |0.4 |- |Trypticase |0.2 |- |Yeast..."

2026-10-22T20:27:39Z

Created page with "== Description == M10 is a medium without rumen fluid for nonselective enumeration and isolation of rumen bacteria [1]. == Recipe == {| class="wikitable" |+ !Component !Percentage in mediuma |- |Glucose |0.05 |- |Cellobiose |0.05 |- |Soluble Starch |0.05 |- |Minerals (v/v)b |3.75 |- |Na2S·9H2O |0.025 |- |Cystein·HCl·H2O |0.025 |- |Resazurin |0.001 |- |Na2CO3 |0.4 |- |Trypticase |0.2 |- |Yeast..."

New page

== Description ==
M10 is a medium without rumen fluid for nonselective enumeration and isolation of rumen bacteria [1].
== Recipe ==
{| class="wikitable"
|+
!Component
!Percentage in mediuma
|-
|Glucose
|0.05
|-
|Cellobiose
|0.05
|-
|Soluble Starch
|0.05
|-
|Minerals (v/v)b
|3.75
|-
|Na2S·9H2O
|0.025
|-
|Cystein·HCl·H2O
|0.025
|-
|Resazurin
|0.001
|-
|Na2CO3
|0.4
|-
|Trypticase
|0.2
|-
|Yeast extract
|0.05
|-
|Volatile fatty acids (v/v)c
|0.31
|-
|Hemin
|0.0001
|}
a Weight/ volume unless otherwise indicated.

b Except for K2HPO4, which was prepared separately, the minerals were prepared as a common stock solution. The final mineral concentrations in the medium were as follows: KH2PO4, 1.3 X 10-3 M; NaCl, 7.6 X 10-4 M; (NH)2SO4, 3.4 X 10-3 M; K2HPO4, 1.7 X 10-3 M; Ca(Cl)2, 4.1 X 10-4 M; MgSO4c·7H20, 3.8 X 10-4 M.

c The volatile fatty acid mixture used was the following: acetic, 17 ml (2.9 X 10-2 M); propionic, 6ml (8.0 X 10-3 M); butyric, 4 ml (4.3 X 10-3 M); isobutyric, 1 ml (1.1 X 10-3 M); n-valeric, isovaleric, and DL-α-methylbutyric, 1 ml each (9 X 10-4 M). The figures in parentheses indicate the final concentration of each acid when 0.31 ml of mixture was placed in a final medium volume of 100 ml.

The final pH of media is 6.7 to 6.8.

=== Directions ===
Suspend 38 g of RCM in 1 liter of distilled water. Boil to dissolve solids completely, aliquot, and sterilize by autoclaving at 121 °C for 15 minutes.

== Notes ==
[[File:Cell pellet formation of Megasphaera elsdenii strain AG22635 in RCM variant.png|thumb|Cell pellet of Megasphaera elsdenii strain AG22635 after a 10-minute 5000 x g centrifuge spin. The culture on the left was grown in commercial RCM. The culture on the right was grown in lab-made RCM without agar.]]
Store bought RCM contains 0.5 g/L agar to mitigate oxygen permeability into the medium. The addition of this agar can have a compounding effect on the formation of [[wikipedia:Extracellular_polymeric_substance|extracellular polymeric substances (EPS)]] in some species like [[Sharpea azabuensis KH1P5|Sharpea azubuensis]] and [[Megasphaera elsdenii T81|Megasphaera elsdeni]]. This EPS generation makes the formation of cell pellets difficult and can affect the procedure of making competent cells. It's recommended to make your RCM "in house" without agar to avoid this complication.

<gallery>
</gallery>

== Sources ==

* [https://documents.thermofisher.com/TFS-Assets/MBD/Specification-Sheets/BT-SPEC-0099.pdf ThermoFisher CM0149] (Used by Guss Lab)